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ID   PACYC177   preliminary; circular DNA; SYN; 3941 BP.
XX
AC   X06402; L08776; ATCC37031;
XX
DT   02-APR-1988 (Rel. 3, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   E. coli plasmid vector pACYC177 - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-3940
RC   pACYC177
RA   Rose R.E.;
RT   ;
RL   Submitted (15-DEC-1987) on tape to the EMBL Data Library by:
RL   Rose R.E., Calgene Inc., 1920 Fifth St., Davis, CA 95616, USA.
XX
RN   [2]
RP   1-3940
RC   pACYC177 from pACYC175
RA   Rose R.E.;
RT   "The nucleotide sequence of pACYC177";
RL   Nucleic Acids Res. 16:356-356(1988).
XX
RN   [3]
RP   1-1226, 1227-1227, 1228-3940
RC   pACYC177
RA   Mok Y.K.;
RT   ;
RL   Submitted (02-MAR-1991) on paper to the EMBL Data Library by:
RL   Mok Y.K., .
XX
RN   [4]
RP   1-3940
RC   from p15A
RC   from pACYC177
RC   from pACYC184
RA   Mok Y.K., Clark D.R., Kam K.M., Shaw P.C.;
RT   "BsiY I, a novel thermophilic restriction endonuclease that
RT   recognizes 5' ccnnnnnnngg 3' and the discovery of a wrongly
RT   sequenced site in pACYC177";
RL   Nucleic Acids Res. 19:2321-2323(1991).
XX
RN   [5]
RC   pACYC139 from p15A & pSC105
RC   pACYC140 from p15A & pSC105
RC   pACYC142 from pACYC140 & pSC204, Tn3
RC   pACYC160 from p15A & pSC101
RC   pACYC161 from pACYC160 & pSC212
RC   pACYC175 from pACYC161 & pKT002
RC   pACYC177 from pACYC175
RC   pACYC184 from pACYC175
RA   Chang A.C., Cohen S.N.;
RT   "Construction and characterization of amplifiable multicopy DNA
RT   cloning vehicles derived from the P15A cryptic miniplasmid";
RL   J. Bacteriol. 134:1141-1156(1978).
XX
RN   [6]
RP   1-3941
RC   pACYC177
RA   Gilbert W.;
RT   "Obtained from VecBase 3.0";
RL   Unpublished (1991).
XX
RN   [7]
RC   pKT007 from R6 & pML21
RC   pKT002 from ColE1 & R6-5
RC   pFC001 from ColE1 & R6-5
RC   pFC012 from ColE1 & R6-5
RA   Timmis K., Cabello F., Cohen S.N.;
RT   "Properties and regulation of DNA replication origins and
RT   associated functions in multi-origin replicons";
RL   Plasmids 0:417-424(1977).
RL   Mitsuhashi S., ed., Avicenum, Prague.
XX
RN   [8]
RC   pSC122 from pSC113
RC   pSC137, pSC138, pSC140, pSC141 from pSC122 & F' lac
RC   pSC135, pSC136, pSC139 from pSC122 & R6-5
RA   Timmis K., Cabello F., Cohen S.N.;
RT   "Cloning, isolation, and characterization of replication regions
RT   of complex plasmid genomes";
RL   Proc. Natl. Acad. Sci. U.S.A. 72:2242-2246(1975).
XX
RN   [9]
RC   from ColE1
RC   from ColE2
RC   from ColV
RC   from ColIb
RA   Helinski D.R., Herschman H.R.;
RT   "Effect of Rec- mutations on the activity of colicinogenic factors";
RL   J. Bacteriol. 94:700-706(1967).
XX
RN   [10]
RC   pSC204 from pSC201 & pSC50, Tn3
RC   pSC301 from pSC101
RC   pSC304 from pSC101 & Tn3
RA   Kretschmer P.J., Cohen S.N.;
RT   "Selected translocation of plasmid genes: frequency and regional
RT   specificity of translocation of the Tn3 element";
RL   J. Bacteriol. 130:888-899(1977).
XX
RN   [11]
RC   pSC211, pSC213, pSC215, pSC217, pSC220 from pSC105 & pSC50, Tn3
RC   pSC221, pSC222 from pSC105 & pSC50, Tn3
RC   PCC10 from pSC105 & Tn21
RC   PCC25 from pSC105 & Tn21
RC   PCC36 from pSC105 & Tn21
RC   pACYC140 from pSC105
RC   [pACYC144 from pSC105]
RC   pSC212, pSC225, pSC226 from pSC211
RC   pSC214, pSC219A, pSC219B, pSC223, pSC224 from pSC213
RC   pSC208 from pSC220
RC   pSC209 from pSC221
RA   Nisen P.D., Kopecko D.J., Chou J., Cohen S.N.;
RT   "DNA sequence deletion at the termini of a transposable
RT   ampicillin-resisatnt element";
RL   J. Mol. Biol. 117:975-998(1977).
XX
RN   [12]
RC   pSC113 from pSC101 & pI258
RA   Chang A.C., Cohen S.N.;
RT   "Genome construction between bacterial species in vitro:
RT   replication and expression of Staphylococcus plasmid genes in
RT   Escherichia coli";
RL   Proc. Natl. Acad. Sci. U.S.A. 71:1030-1034(1974).
XX
CC   The sequence is circular; however, the sequence starts with a HincII
CC   site.
CC   Bases 564-841, 1494-1721, and 2869-3039 were determined
CC   while the rest of the sequence was taken from the published
CC   sequences of Tn903, Tn3, and p15A.
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pACYC177)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (NEB)(ATCC)
CC   HO (E.coli 1200)(E.coli C600 chi1776)(E.coli)
CC   CP (low)
CC   FN (cloning)
CC   SE ()
CC   PA (Tn3)(Tn903)(p15A)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pSC101 EcoRI 9263bp 9262..9262
FT                   2. R6-5 EcoRI-EcoRI 6900bp, Tn903 kan gene/#V00359
FT                   -> pSC105 16163bp
FT                   1. p15A HincII
FT                   2. pSC105 EcoRI-SalI, Tn903 kan gene/#V00359
FT                   reverse transcriptase fill in:reverse transcriptase
FT                   \ fill in
FT                   -> pACYC140 4600bp
FT                   1. R1
FT                   -> R1-19
FT                   1. R1-19, Tn3/Tn4 amp gene
FT                   -> pSC50
FT                   1. pSC105
FT                   -> pSC205
FT                   1. pSC205
FT                   -> pSC201
FT                   1. pSC201
FT                   2. pSC50, Tn3 amp gene
FT                   -> pSC204
FT                   1. pACYC140
FT                   transposition
FT                   2. pSC204, Tn3 amp gene
FT                   -> pACYC142 10800bp
FT                   1. pACYC142 remove HaeII-HaeII fragments and religate
FT                   -> pACYC177 3941bp [amp, kan]"
FT   misc_feature    complement(3698..3940)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp), from Tn3 [2]"
FT   misc_feature    complement(1..615)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp), from Tn3 [2]"
FT   misc_binding    1..1
FT                   /note="SIT unique HincII"
FT   misc_binding    18..18
FT                   /note="SIT unique BcgI"
FT   misc_binding    62..62
FT                   /note="SIT unique ScaI"
FT   misc_binding    299..299
FT                   /note="SIT unique PstI"
FT   misc_binding    320..320
FT                   /note="SIT unique FspI"
FT   misc_binding    421..421
FT                   /note="SIT unique BglI"
FT   misc_binding    455..455
FT                   /note="SIT unique BpmI"
FT   misc_binding    473..473
FT                   /note="SIT unique BsaI"
FT   misc_binding    524..524
FT                   /note="SIT unique Pfl108I"
FT   misc_binding    540..540
FT                   /note="SIT unique Eam1105I"
FT   misc_binding    592..592
FT                   /note="SIT unique BanI"
FT   misc_feature    complement(615..3698)
FT                   /note="TRN Tn3 with beta-lactamase gene"
FT   repeat_unit     728..765
FT                   /note="TRN Tn3 inverted repeat"
FT   misc_feature    complement(756..3026)
FT                   /note="TRN Tn3 fragment [2]"
FT   rep_origin      766..1595
FT                   /note="ORI E. coli miniplasmid p15A [5]"
FT   misc_binding    987..987
FT                   /note="SIT unique AlwNI"
FT   misc_binding    1013..1013
FT                   /note="SIT unique NspI"
FT   misc_binding    1096..1096
FT                   /note="SIT unique BspGI"
FT   conflict        1226..1228
FT                   /note="replace(1226..1228,ag); ref. [1] & [2]"
FT   rep_origin      1326..1328
FT                   /note="ORI E. coli miniplasmid p15A (start) [5]"
FT   misc_binding    1338..1338
FT                   /note="SIT unique SacII"
FT   misc_binding    1500..1500
FT                   /note="SIT unique SgrAI"
FT   misc_binding    1505..1505
FT                   /note="SIT unique BsgI"
FT   misc_binding    1576..1576
FT                   /note="SIT unique AccI"
FT   misc_binding    1587..1587
FT                   /note="SIT unique NheI"
FT   repeat_unit     1590..1817
FT                   /note="TRN Tn903 inverted repeat"
FT   misc_feature    1595..3025
FT                   /note="TRN HaeII-HaeII fragment from Tn903 [2]"
FT   misc_binding    1817..1817
FT                   /note="SIT unique DraIII"
FT   misc_feature    1922..2734
FT                   /note="ANT E. coli neomycin phosphotransferase gene
FT                   (NPT), neomycin resistance gene (neo)
FT                   kanamycin resistance gene (kan), from Tn3 [2]"
FT   misc_binding    1951..1951
FT                   /note="SIT unique XhoI"
FT   misc_binding    2004..2004
FT                   /note="SIT unique BanII"
FT   misc_binding    2008..2008
FT                   /note="SIT unique NruI"
FT   misc_binding    2043..2043
FT                   /note="SIT unique ClaI"
FT   misc_binding    2225..2225
FT                   /note="SIT unique SmaI"
FT   misc_binding    2261..2261
FT                   /note="SIT unique EcoNI"
FT   misc_binding    2363..2363
FT                   /note="SIT unique Esp3I"
FT   misc_binding    2471..2471
FT                   /note="SIT unique HindIII"
FT   misc_binding    2610..2610
FT                   /note="SIT unique PflMI"
FT   misc_binding    2790..2790
FT                   /note="SIT unique BsmFI"
FT   repeat_unit     2798..3025
FT                   /note="TRN Tn903 inverted repeat"
FT   misc_feature    3025..3515
FT                   /note="TRN Tn3 repressor fragment"
FT   misc_binding    3278..3278
FT                   /note="SIT unique BstEII"
FT   misc_binding    3319..3319
FT                   /note="SIT unique BamHI"
FT   misc_binding    3396..3396
FT                   /note="SIT unique DrdI"
FT   misc_binding    3469..3469
FT                   /note="SIT unique FauI"
FT   misc_binding    3495..3495
FT                   /note="SIT unique BbsI"
FT   misc_binding    3504..3504
FT                   /note="SIT unique EcoO109I"
FT   misc_binding    3562..3562
FT                   /note="SIT unique AatII"
FT   misc_binding    3798..3798
FT                   /note="SIT unique Eco57I"
FT   misc_binding    3813..3813
FT                   /note="SIT unique ApaLI"
XX
SQ   Sequence 3941 BP; 1026 A; 893 C; 993 G; 1029 T; 0 other;
     gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccttaat aagatgatct
     tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     ccaacatagt aagccagtat acactccgct agcgctgagg tctgcctcgt gaagaaggtg
     ttgctgactc ataccaggcc tgaatcgccc catcatccag ccagaaagtg agggagccac
     ggttgatgag agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca
     cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc
     gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata
     aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg
     atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     aacactggca gagcattacg ctgacttgac gggacggcgg ctttgttgaa taaatcgaac
     ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     ctccctcact ttctggctgg atgatggggc gattcaggcc tggtatgagt cagcaacacc
     ttcttcacga ggcagacctc agcgctcaaa gatgcagggg taaaagctaa ccgcatcttt
     accgacaagg catccggcag ttcaacagat cgggaagggc tggatttgct gaggatgaag
     gtggaggaag gtgatgtcat tctggtgaag aagctcgacc gtcttggccg cgacaccgcc
     gacatgatcc aactgataaa agagtttgat gctcagggtg tagcggttcg gtttattgac
     gacgggatca gtaccgacgg tgatatgggg caaatggtgg tcaccatcct gtcggctgtg
     gcacaggctg aacgccggag gatcctagag cgcacgaatg agggccgaca ggaagcaaag
     ctgaaaggaa tcaaatttgg ccgcaggcgt accgtggaca ggaacgtcgt gctgacgctt
     catcagaagg gcactggtgc aacggaaatt gctcatcagc tcagtattgc ccgctccacg
     gtttataaaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     gcacttttaa agttctgcta tgtggcgcgg tattatcccg t
//