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ID   PAM35      preliminary; circular DNA; SYN; 4805 BP.
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AC   M69063; X07465; X04395; ATCC77186;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pAM35 - complete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RC   lacZPO:RNAI from pUC9 & pUC18 & pGB2
RC   pAM22 from pBR322 & pAM1
RC   pAM28 from pAM22 & lacZPO:RNAI
RC   aadA-cassette from pCK1 & pCKT & pSB118
RC   pAM34 from aadA-cassette & pAM28
RC   pAM35 from aadA-cassette & pAM28, no lacIq gene
RC   pAM39 from pOM41 & pAM34
RA   Gil D., Bouche J.P.;
RT   "ColE1-type vectors with fully repressible replication";
RL   Gene 105:17-22(1991).
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RN   [2]
RC   pJPB19 from pBR322
RC   pJPB38 from pBS28 & pBS29 & pHP45omega
RC   pKC1 from pGB2 & E.coli chromosome
RC   pKC7 from pUC9 & E.coli chromosome
RC   pKC8 from pUC8 & E.coli chromosome
RC   pKC17 from pKC7
RC   pKC18 from pKC8
RC   pKC27 from pGEM-1 & E.coli chromosome
RC   pKC28 from pGEM-1 & E.coli chromosome
RC   pKC36 from pGEM-2 & E.coli chromosome
RC   pKC37 from pGEM-1 & E.coli chromosome
RC   pJPB48 from pGEM-1 & E.coli chromosome
RC   pJPB50 from pMC1403 & E.coli chromosome
RA   Cam K., Bejar S., Gil D., Bouche J.P.;
RT   "Identification and sequence of gene dicB: translation of the
RT   division inhibitor from an in-phase internal start";
RL   Nucleic Acids Res. 16:6327-6338(1988).
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RN   [3]
RC   lambda JS1 from lambda & E.coli chromosome Hfr KL99
RC   pBS27 from pHC79 & lambda JS1
RC   pBS28 from pBS27
RC   pBS28omega4 from pBS28 & omega, spec gene
RC   pBS29 from pBS28
RC   pBS32 from pBS29
RC   pBS37 from pBS27 & pBR322
RC   pJPB36 from pUC9 & pBS28omega4
RC   pBS44 from pJPB36
RA   Bejar S., Bouche J.P.;
RT   "A new dispensable genetic locus of the terminus region involved
RT   in control of cell division in Escherichia coli";
RL   Mol. Gen. Genet. 201:146-150(1985).
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RN   [4]
RC   pBS39 from pBR322 & E.coli chromosome & pHP45omega
RA   Bejar S., Cam K., Bouche J.P.;
RT   "Control of cell division in Escherichia coli. DNA sequence of
RT   dicA and of a second gene complementing mutation dicA1, dicC";
RL   Nucleic Acids Res. 14:6821-6833(1986).
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RN   [5]
RC   pJPB3 from Tn10
RA   Bouche J.P., Gelunge J.P., Louarn J., Louarn J.M., Kaiser K.;
RT   "Relationships between th ephysical and genetic maps of a 470
RT   kilobase pair region around the terminus of Escherichia coli K12
RT   DNA replication";
RL   J. Mol. Biol. 154:21-32(1982).
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RN   [6]
RC   pK5 from pBR322 & E.coli chromosome, Tn10 tet gene
RA   Kaiser K.;
RT   "The origin of Q-independent derivatives of phage lambda";
RL   Mol. Gen. Genet. 179:547-554(1980).
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CC   Created by Moore, July 1995, under contract with NCBI.
CC   Integration vector permitting positive selection for inserts.
CC   It can be propagated in a lacI+ strain such as C600 in the absence of
CC   IPTG, but replication is inefficient. [1]
CC   Transcription of the replication primer RNA is regulated by the lacZpo
CC   promoter/operator.  In the presence of lacIq, replication is entirely
CC   dependent on the presence of inducer. [1]
CC   The repressible replication system permits temporary maintenance of
CC   the plasmid, construction of strains with stable integrants of vector
CC   derivatives, and recovery of sequences adjacent to cloned fragments.
CC   Can be selected with 100 ug/ml ampicillin and/or 75 ug/ml
CC   spectinomycin.
CC   Permits positive selection because deletion of the aadA fragment is
CC   not viable. [1]
CC   pAM35 (ATCC 77186) differs from pAM34 (ATCC 77185) by deletion of the
CC   DraII fragment in pAM34 carrying lacIq. [1]
CC   E. coli JS219 is MC1061 malPp delta534::lacIq. [2]
CC   The order of the major features in this plasmid is: lacZpo - ori - rom
CC   - transcription terminator - BamHI - MCS - XmaIII - aadA (spcR) -
CC   HindIII - MCS - BamHI - transcription terminator - bla. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--4.8; BamHI--3.0, 1.8; KpnI--3.4, 1.4. (ATCC staff)
CC   Medium is -1 1227 + IPTG (1 mM).
CC   NM (pAM35)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli JS219)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pAM34)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pGB series
FT                   -> pGB2
FT                   1. pUC9 HinP1I/HhaI-HindIII 181bp 53..234,
FT                   \ lacZPO promoter
FT                   2. pUC9 Fnu4HI/BsoFI/ItaI-MaeI 121bp 1926..2047, RNAI
FT                   3. pUC18, MCS
FT                   4. pGB2
FT                   -> lacZPO:RNAI
FT                   1. E. coli, lacZ/lacPO/lacI gene
FT                   -> pAM1
FT                   1. pBR322 AvaI-EcoRI 2934bp 1426..4360, ori/amp
FT                   S1 nuclease
FT                   Klenow:Klenow
FT                   2. pAM1 HindIII-EcoRI 1200bp, lacI gene
FT                   -> pAM22 4100bp
FT                   1. pAM22 MaeI 4100bp, pBR322 1490..1490
FT                   \ between ori and lacI gene
FT                   2. lacZPO:RNAI MaeI-MaeI, lacZPO/RNAI gene
FT                   -> pAM28 4101bp
FT                   1. pUC18 2686bp, MCS
FT                   2. oligo
FT                   -> pSB118 2700bp
FT                   1. phage T4 gene 32
FT                   2. R100.1, omega/aadA gene
FT                   -> pCK1
FT                   1. phage T4 gene 32
FT                   2. R100.1, omega/aadA gene
FT                   -> pCK2
FT                   1. phage T4 gene 32
FT                   2. pCK2
FT                   -> pCKT
FT                   1. pCK1, aadA gene
FT                   2. pCKT, phage T4 gene 32 terminators
FT                   3. pSB118 2700bp, MCS
FT                   -> aadA-cassette
FT                   1. aadA-cassette DraI-DraI 1900bp, MCS-aadA-MCS
FT                   2. pAM28 remove DraI-DraI 19bp, pBR322 3235..3254
FT                   \ no lacIq gene/4082bp
FT                   -> plasmid 6000bp
FT                   1. plasmid PstI 6000bp, pBR322 3614..3614
FT                   fill in
FT                   -> pAM34 6000bp
FT                   1. pAM34 remove DraII/EcoO109I-DraII/EcoO109I 1195bp
FT                   \ 4757..5952, lacIq gene/4805bp
FT                   \ DraII = 1798 1808 3601 3611 4757 5952 5994
FT                   -> pAM35 4805bp"
FT   -               1..4756
FT                   /note="pAM34 1..4756 4756bp
FT                   DraII = RG^GNCCY"
FT   -               4757..4805
FT                   /note="pAM34 5952..6000 49bp"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   misc_binding    0..0
FT                   /note="MCS BamHI-SalI-PstI-SphI-XmaIII"
FT   CDS             0..0
FT                   /note="ANT E. coli spectinomycin resistance gene
FT                   (spc)"
FT   terminator      0..0
FT                   /note="GEN E. coli T4 gene 32 terminator"
FT   misc_binding    0..0
FT                   /note="MCS HindIII-EcoRI-SacI-KpnI-BamHI"
FT   misc_binding    0..0
FT                   /note="SIT unique SalI-PstI-SphI-XmaIII-
FT                   HindIII-EcoRI-SacI-KpnI"
FT   CDS             complement(0..0)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (amp/apr)"
FT   CDS             complement(0..0)
FT                   /note="REP E. coli lacIq repressor gene"
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SQ   Sequence 4805 BP; 1189 A; 1208 C; 1237 G; 1171 T; 0 other;
     ctagagtcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     aaacaggggt accctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     cgtgtaaggg ggaattctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagatc
     cttttagctt tatgcttgta aaccgttttg tgaaaaaatt tttaaaataa aaaaggggac
     ctctagggtc cccaattaat tagtaatata atctattaaa ggtcattcaa aaggtcatcc
     accggatcct ctagagtcga cctgcaggca tgcccgttcc atacagaagc tgggcgaaca
     aacgatgctc gccttccaga aaaccgagga tgcgaaccac ttcatccggg gtcagcacca
     ccggcaagcg ccgcgacggc cgaggtcttc cgatctcctg aagccagggc agatccgtgc
     acagcacctt gccgtagaag aacagcaagg ccgccaatgc ctgacgatgc gtggagaccg
     aaaccttgcg ctcgttcgcc agccaggaca gaaatgcctc gacttcgctg ctgcccaagg
     ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     tcaccaaggt agtcggcaaa taatgtctaa caattcgttc aagccgacgc cgcttcgcgg
     cgcggcttaa ctcaagcgtt agatgcacta agcacataat tgctcacagc caaactatca
     ggtcaagtct gcttttatta tttttaagcg tgcataataa gccctacaca aattgggaga
     tatatcatga aaggctggct ttttcttgtt atcgcaatag ttggcgaagt aatcgcaaca
     tccgcattaa aatctagcga gggctttact aagcttatcg aattcgagct cggtaccccg
     gggatccggt ggatgacctt ttgaatgacc tttaatagat tatattacta attaattggg
     gaccctagag gtcccctttt ttattttaaa aattttttca caaaacggtt tacaagcata
     aagctaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     ataggcgtat cacgaggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga
     cccgg
//