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ID   2MICRONA   preliminary; circular DNA; FUN; 6318 BP.
XX
AC   J01347; L00321; L00322; L00323; L00324; M10185; M11111; M11593;
AC   M14239; M14240; M14241; M14242; M14243; M14244; M14245; M14253;
AC   M14254; M14255; M14256; M14257; M14258; M14259; M14591; M14592;
AC   M14593; M14594; M14595; M14596; M14597; M14598; V01323;
XX
DT   01-JUL-1994 (Rel. 9, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   Saccharomyces plasmid 2 micron A form - complete.
XX
KW   cloning vector.
XX
OS   Saccharomyces cerevisiae (yeast)
OC   Eukaryota;; Fungi; Eumycota; Hemiascomycetes; Endomycetales;
OC   Saccharomycetaceae.
XX
RN   [1]
RP   1-1022
RC   pJDB71 from pMB9 & yeast 2 micron form A
RA   Hindley J., Phear G.A.;
RT   "Sequence of 1019 nucleotides encompassing one of the inverted
RT   repeats from the yeast 2 micron plasmid";
RL   Nucleic Acids Res. 7:361-375(1979).
XX
RN   [2]
RP   1-6318
RC   p82-6B from pMB9 & yeast 2 micron
RA   Hartley J.L., Donelson J.E.;
RT   "Nucleotide sequence of the yeast plasmid";
RL   Nature 286:860-865(1980).
XX
RN   [3]
RP   3891-3990
RC   from yeast 2 micron, recombination site
RA   Broach J.R., Guarascio V.R., Jayaram M.;
RT   "Recombination within the yeast plasmid 2-micron circle is
RT   site-specific";
RL   Cell 29:227-234(1982).
XX
RN   [4]
RP   3881-4020
RC   pMMD2 from pBR322 & yeast LEU2 gene & yeast URA3 gene & yeast 2 micron
RA   McLeod M., Volkert F., Broach J.R.;
RT   "Components of the site-specific recombination system encoded by the
RT   yeast plasmid 2-micron circle";
RL   Cold Spring Harb. Symp. Quant. Biol. 49:779-787(1984).
XX
RN   [5]
RP   670-732
RC   pBA112 from pUC9 & yeast FLP recombination site
RA   Andrews B.J., Proteau G.A., Beatty L.G., Sadowski P.D.;
RT   "The FLP recombinase of the 2 micron circle DNA of yeast:
RT   interaction with its target sequences";
RL   Cell 40:795-803(1985).
XX
RN   [6]
RP   5570-5605
RC   plasmid from yeast 2 micron FLP & pMA301
RC   pDV64 from plasmid & pDR540
RC   pBA101 from pCV19 & pBR322
RC   pBA104 from pBA101 & pCV19
RC   pBA107 from pBA101 & pCV19
RA   Babineau D., Vetter D., Andrews B.J., Gronostajski R.M.,
RA   Proteau G.A., Beatty L.G., Sadowski P.D.;
RT   "The FLP protein of the 2-micron plasmid of yeast: purification of
RT   the protein from Escherichia coli cells expressing the cloned FLP
RT   gene";
RL   J. Biol. Chem. 260:12313-12319(1985).
XX
RN   [7]
RC   plasmid from pUC9 & yeast 2 micron, FLP cleavage site
RC   pGP40 from plasmid & yeast 2 micron
RA   Gronostajski R.M., Sadowski P.D.;
RT   "Determination of DNA sequences essential for FLP-mediated
RT   recombination by a novel method";
RL   J. Biol. Chem. 260:12320-12327(1985).
XX
RN   [8]
RC   pBR2micronA from pBR322 & yeast 2 micron A form
RC   from yeast 2 micron, CAP site/polyA
RA   Sutton A., Broach J.R.;
RT   "Signals for transcription initiation and termination in the
RT   Saccharomyces cerevisiae plasmid 2 micron circle";
RL   Mol. Cell. Biol. 5:2770-2780(1985).
XX
RN   [9]
RC   from pBA112, FLP recombination site
RA   Gronostajski R.M., Sadowski P.D.;
RT   "The FLP recombinase of the Saccharomyces cerevisiae 2-micron
RT   plasmid attaches covalently to DNA via a phosphotyrosyl linkage";
RL   Mol. Cell. Biol. 5:3274-3279(1985).
XX
RN   [10]
RP   667-739
RC   pJFS166 from pXf3 & linker & yeast 2 micron, FLP binding site
RC   pJFS122, pJFS110, pJFS105, pJFS100 from pJFS166 & M13mp8
RC   pJFS30 series from pJFS100
RA   Senecoff J.F., Bruckner R.C., Cox M.M.;
RT   "The FLP recombinase of the yeast 2-micron-m plasmid:
RT   Characterization of its recombination site";
RL   Proc. Natl. Acad. Sci. U.S.A. 82:7270-7274(1985).
XX
RN   [11]
RC   pWP series from pMMD2 & linker
RC   pMu5T2-2, p545-2, p624B-2, p613B-2 from pMMD2
RC   pINV1 from pMMD2 & pCV20
RA   McLeod M., Craft S., Broach J.R.;
RT   "Identification of the crossover site during FLP-mediated
RT   recombination in the Saccharomyces cerevisiae plasmid 2 micron
RT   circle";
RL   Mol. Cell. Biol. 6:3357-3367(1986).
XX
RN   [12]
RP   1-6318
RC   yeast 2 micron
RA   Senecoff J.;
RT   ;
RL   Submitted (24-JAN-1986) by:
RL   Senecoff J., .
XX
RN   [13]
RC   from yeast 2 micron
RC   from yeast 2 micron & pBR322 & LEU2 gene
RA   Broach J.R., Hicks J.B.;
RT   "Replication and recombination functions associated with the yeast
RT   2micron circle";
RL   Cell 21:501-508(1980).
XX
RN   [14]
RC   from yeast 2 micron
RA   Broach J.R.;
RT   "The yeast plasmid 2 micron circle";
RL   Cell 28:203-204(1982).
XX
RN   [15]
RC   pMA301 from YEp series
RA   Kingsman S., Kingsman A.;
RT   ;
RL   Unpublished (1985).
XX
RN   [16]
RC   pCV20 or pMJ5 from pBR322 & yeast 2 micron & LEU2 gene
RC   pXho series from pCV20
RA   Jayaram M., Li Y.Y., Broach J.R.;
RT   "The yeast plasmid 2 micron circle encodes components required for
RT   its high copy propagation";
RL   Cell 34:95-104(1983).
XX
CC   Yeast 2 micron plasmid contains two 599 bp inverted repeats
CC   separated by a large unique (UL) and a small unique (US) region.
CC   During recombination the UL and US regions invert producing two
CC   sequence forms that differ in the orientation of one unique region
CC   relative to the other.  The A form is presented below.
CC   FLP is the only 2-micron circle-encoded protein needed for specific
CC   site recombination between the IRs of 2-micron circle.  The minimal
CC   size of the recombination site required for efficient FLP
CC   recombinase-catalyzed recombination in vitro is no more than 28 bp,
CC   which includes parts of two 13 bp inverted repeats (positions
CC   690-702 and 711-723) and all of an 8 bp spacer (703-710) [5].  The
CC   FLP recombinase cleaves the DNA at the boundaries of the spacer and
CC   becomes covalently linked to the spacer DNA [5],[9].  The
CC   efficiency of the recombination is reduced if the spacer in a
CC   recombinant site is increased or decreased by 1 bp, while the
CC   spacer in the second site is unaltered [5].  Recombination between
CC   two sites with identical 1-base pair additions or deletions is
CC   relatively unaffected, suggesting that pairing of sequences in the
CC   spacer regions is important in FLP-promoted recombination events
CC   [5].  The sequence asymmetry utilized by the recombinase to
CC   determine the orientation of the site is located uniquely within
CC   the spacer region. Another 13 bp direct repeat, is found at
CC   positions 676-688 [5].  FLP-mediated recombination involving two
CC   FLP sites that are inverted with respect to each other results in
CC   inversion of the DNA sequences between the sites [4].  If the
CC   participating recombination sites are in direct orientation, FLP
CC   promotes only the excision of the intervening DNA sequences [4].
CC   The Rep 1 and Rep proteins are involved plasmid partitioning and
CC   protein stability.
CC   A start codon in phase with the Rep1 coding region is located at
CC   positions 1966-1964.  Two CAP sites for Rep1 mRNA are located
CC   beyond the 'atg' codon (position 2008) at positions 2004 and 2005.
CC   Complete source information:
CC   Yeast (S.cerevisiae, strain A364A D5) DNA, clones pJDB71 [1],
CC   p82-6B [2], pCV20 [3], pMMD2 [4], pGP20 [5], pJFS166 [10].
CC   NCBI gi: 172190 & 4182
CC   NM (yeast 2 micron A form)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (S.cerevisiae)
CC   CP (high)
CC   FN (cloning)
CC   SE ()
CC   PA (2 micron)
CC   BR ()
CC   OF (YEp)(YIp)(2micron Circle-pBR322-LEU2)
CC   OR (1 bp upstream of EcoRI site)
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. S. cerevisiae
FT                   -> yeast 2 micron 6318bp"
FT   mRNA            join(5549..6318,1..545)
FT                   /note="Flp mRNA"
FT   repeat_region   341..939
FT                   /note="IR2"
FT   misc_binding    673..722
FT                   /note="FLP recombinase binding site A [9]"
FT   misc_recomb     702..703
FT                   /note="US end/UL start [9],[7]"
FT   mRNA            complement(836..2004)
FT                   /note="Rep1 mRNA (alt.)"
FT   mRNA            complement(836..2005)
FT                   /note="Rep1 mRNA (alt.)"
FT   mRNA            complement(836..2010)
FT                   /note="Rep1 mRNA (alt.)"
FT   mRNA            complement(836..2017)
FT                   /note="Rep1 mRNA (alt.)"
FT   mRNA            complement(836..2019)
FT                   /note="Rep1 mRNA (alt.)"
FT   mRNA            complement(836..2038)
FT                   /note="Rep1 mRNA (alt.)"
FT   mRNA            2254..2841
FT                   /note="D mRNA (alt.; 5' end +/- 3 bp)"
FT   mRNA            2254..2861
FT                   /note="D mRNA (alt.; 5' end +/- 3 bp)"
FT   repeat_region   3714..4312
FT                   /note="IR1"
FT   misc_binding    3930..3979
FT                   /note="FLP-recombinase binding site B [9]"
FT   misc_recomb     3941..3942
FT                   /note="UL end/US start [9],[7]"
FT   mRNA            complement(4108..5184)
FT                   /note="REP2 mRNA (major alt.)"
FT   mRNA            complement(4108..5183)
FT                   /note="REP2 mRNA (major alt.)"
FT   mRNA            complement(4108..5182)
FT                   /note="REP2 mRNA (major alt.)"
FT   mRNA            complement(4108..5198)
FT                   /note="REP2 mRNA (major alt.)"
FT   mRNA            complement(4108..5197)
FT                   /note="REP2 mRNA (major alt.)"
FT   mRNA            complement(4108..5196)
FT                   /note="REP2 mRNA (major alt.)"
FT   mRNA            complement(4108..5195)
FT                   /note="REP2 mRNA (major alt.)"
FT   mRNA            complement(4108..5212)
FT                   /note="REP2 mRNA (minor alt.)"
FT   mRNA            complement(4108..5223)
FT                   /note="REP2 mRNA (minor alt.)"
FT   CDS             join(5570..6318,1..523)
FT                   /note="recombinase;  (FLP);  NCBI gi: 172191."
FT   CDS             complement(887..2008)
FT                   /note="Rep 1 protein;  NCBI gi: 172192."
FT   CDS             2271..2816
FT                   /note="D protein;  NCBI gi: 172193."
FT   CDS             complement(4308..5198)
FT                   /note="Rep 2 protein;  NCBI gi: 172194."
XX
SQ   Sequence 6318 BP; 1876 A; 1284 C; 1179 G; 1979 T; 0 other;
     gaattctgaa ccagtcctaa aacgagtaaa taggaccggc aattcttcaa gcaataaaca
     ggaataccaa ttattaaaag ataacttagt cagatcgtac aataaagctt tgaagaaaaa
     tgcgccttat tcaatctttg ctataaaaaa tggcccaaaa tctcacattg gaagacattt
     gatgacctca tttctttcaa tgaagggcct aacggagttg actaatgttg tgggaaattg
     gagcgataag cgtgcttctg ccgtggccag gacaacgtat actcatcaga taacagcaat
     acctgatcac tacttcgcac tagtttctcg gtactatgca tatgatccaa tatcaaagga
     aatgatagca ttgaaggatg agactaatcc aattgaggag tggcagcata tagaacagct
     aaagggtagt gctgaaggaa gcatacgata ccccgcatgg aatgggataa tatcacagga
     ggtactagac tacctttcat cctacataaa tagacgcata taagtacgca tttaagcata
     aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg
     tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt
     tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac
     ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
     gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
     aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
     ttcgctcctt gaacttgcat ctaaactcga cctctacatc aacaggcttc caatgctctt
     caaattttac tgtcaagtag acccatacgg ctgtaatatg ctgctcttca taatgtaagc
     ttatctttat cgaatcgtgt gaaaaactac taccgcgata aacctttacg gttccctgag
     attgaattag ttcctttagt atatgataca agacactttt gaactttgta cgacgaattt
     tgaggttcgc catcctctgg ctatttccaa ttatcctgtc ggctattatc tccgcctcag
     tttgatcttc cgcttcagac tgccattttt cacataatga atctatttca ccccacaatc
     cttcatccgc ctccgcatct tgttccgtta aactattgac ttcatgttgt acattgttta
     gttcacgaga agggtcctct tcaggcggta gctcctgatc tcctatatga cctttatcct
     gttctctttc cacaaactta gaaatgtatt catgaattat ggagcaccta ataacattct
     tcaaggcgga gaagtttggg ccagatgccc aatatgcttg acatgaaaac gtgagaatga
     atttagtatt attgtgatat tctgaggcaa ttttattata atctcgaaga taagagaaga
     atgcagtgac ctttgtattg acaaatggag attccatgta tctaaaaaat acgcctttag
     gccttctgat accctttccc ctgcggttta gcgtgccttt tacattaata tctaaaccct
     ctccgatggt ggcctttaac tgactaataa atgcaaccga tataaactgt gataattctg
     ggtgatttat gattcgatcg acaattgtat tgtacactag tgcaggatca ggccaatcca
     gttctttttc aattaccggt gtgtcgtctg tattcagtac atgtccaaca aatgcaaatg
     ctaacgtttt gtatttctta taattgtcag gaactggaaa agtccccctt gtcgtctcga
     ttacacacct actttcatcg tacaccatag gttggaagtg ctgcataata cattgcttaa
     tacaagcaag cagtctctcg ccattcatat ttcagttatt ttccattaca gctgatgtca
     ttgtatatca gcgctgtaaa aatctatctg ttacagaagg ttttcgcggt ttttataaac
     aaaactttcg ttacgaaatc gagcaatcac cccagctgcg tatttggaaa ttcgggaaaa
     agtagagcaa cgcgagttgc attttttaca ccataatgca tgattaactt cgagaaggga
     ttaaggctaa tttcactagt atgtttcaaa aacctcaatc tgtccattga atgccttata
     aaacagctat agattgcata gaagagttag ctactcaatg ctttttgtca aagcttactg
     atgatgatgt gtctactttc aggcgggtct gtagtaagga gaatgacatt ataaagctgg
     cacttagaat tccacggact atagactata ctagtatact ccgtctactg tacgatacac
     ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc
     cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta
     gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat
     ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta
     atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg
     aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat
     atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg
     catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc
     ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa
     aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
     acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc
     agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac
     ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
     ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
     atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
     tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
     ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
     ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca taccctagaa
     gtattacgtg attttctgcc ccttaccctc gttgctactc tccttttttt cgtgggaacc
     gctttagggc cctcagtgat ggtgttttgt aatttatatg ctcctcttgc atttgtgtct
     ctacttcttg ttcgcctgga gggaacttct tcatttgtat tagcatggtt cacttcagtc
     cttccttcca actcactctt tttttgctgt aaacgattct ctgccgccag ttcattgaaa
     ctattgaata tatcctttag agattccggg atgaataaat cacctattaa agcagcttga
     cgatctggtg gaactaaagt aagcaattgg gtaacgacgc ttacgagctt cataacatct
     tcttccgttg gagctggtgg gactaataac tgtgtacaat ccatttttct catgagcatt
     tcggtagctc tcttcttgtc tttctcgggc aatcttccta ttattatagc aatagatttg
     tatagttgct ttctattgtc taacagcttg ttattctgta gcatcaaatc tatggcagcc
     tgacttgctt cttgtgaaga gagcatacca tttccaatcg aatcaaacct ttccttaacc
     atcttcgcag caggcaaaat tacctcagca ctggagtcag aagatacgct ggaatcttct
     gcgctagaat caagaccata cggcctaccg gttgtgagag attccatggg ccttatgaca
     tatcctggaa agagtagctc atcagactta cgtttactct ctatatcaat atctacatca
     ggagcaatca tttcaataaa cagccgacat acatcccaga cgctataagc tgtacgtgct
     tttaccgtca gattcttggc tgtttcaatg tcgtccattt tggttttctt ttaccagtat
     tgttcgtttg ataatgtatt cttgcttatt acattataaa atctgtgcag atcacatgtc
     aaaacaactt tttatcacaa gatagtaccg caaaacgaac ctgcgggccg tctaaaaatt
     aaggaaaagc agcaaaggtg catttttaaa atatgaaatg aagataccgc agtaccaatt
     attttcgcag tacaaataat gcgcggccgg tgcatttttc gaaagaacgc gagacaaaca
     ggacaattaa agttagtttt tcgagttagc gtgtttgaat actgcaagat acaagataaa
     tagagtagtt gaaactagat atcaattgca cacaagatcg gcgctaagca tgccacaatt
     tggtatatta tgtaaaacac cacctaaggt gcttgttcgt cagtttgtgg aaaggtttga
     aagaccttca ggtgagaaaa tagcattatg tgctgctgaa ctaacctatt tatgttggat
     gattacacat aacggaacag caatcaagag agccacattc atgagctata atactatcat
     aagcaattcg ctgagtttcg atattgtcaa taaatcactc cagtttaaat acaagacgca
     aaaagcaaca attctggaag cctcattaaa gaaattgatt cctgcttggg aatttacaat
     tattccttac tatggacaaa aacatcaatc tgatatcact gatattgtaa gtagtttgca
     attacagttc gaatcatcgg aagaagcaga taagggaaat agccacagta aaaaaatgct
     taaagcactt ctaagtgagg gtgaaagcat ctgggagatc actgagaaaa tactaaattc
     gtttgagtat acttcgagat ttacaaaaac aaaaacttta taccaattcc tcttcctagc
     tactttcatc aattgtggaa gattcagcga tattaagaac gttgatccga aatcatttaa
     attagtccaa aataagtatc tgggagtaat aatccagtgt ttagtgacag agacaaagac
     aagcgttagt aggcacatat acttctttag cgcaaggggt aggatcgatc cacttgtata
     tttggatgaa tttttgag
//