back Return to this vector's summary.
ID   PAM34      preliminary; circular DNA; SYN; 6000 BP.
XX
AC   M69063; ATCC77185;
XX
DT   28-MAY-1992 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   E. coli plasmid vector pAM34 - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-6000
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).
XX
CC   Permits positive selection because deletion of the aadA fragment is
CC   not viable.
CC   pAM35 (ATCC 77186) differs from pAM34 (ATCC 77185) by deletion of
CC   the DraII fragment in pAM34 carrying lacIq.
CC   The order of the major features in this plasmid is: lacZpo - ori -
CC   rom - transcription terminator- BamHI - MCS - XmaIII - aadA (spcR) -
CC   MCS - BamHI - transcription terminator - bla - lacIq. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--6.0; BamHI--4.3,1.8; HindIII--6.0; PstI--6.0; KpnI--3.4, 2.7.
CC   (ATCC staff)
CC   Medium is -1 1227 + IPTG (1 mM).
CC   NM (pAM34)
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 ()
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC9 HindIII-HhaI/HinP1I 183bp 234..407,
FT                   \ lacZPO promoter [181bp]
FT                   2. pUC9 Fnu4HI-MaeI 121bp, RNAI gene
FT                   \ MaeI = 1459 1794 2047
FT                   3. pUC18
FT                   4. pGB2
FT                   -> lacZPO:RNAI
FT                   1. pBR322 EcoRI-AvaI 2934bp 1426..4360, ori/amp gene
FT                   S1 nuclease
FT                   Klenow:Klenow
FT                   2. pAM1 HindIII-EcoRI, lacI gene
FT                   -> pAM22 4100bp
FT                   1. pAM22 MaeI, 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
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 2686bp, MCS
FT                   -> aadA-cassette 6000bp
FT                   1. aadA-cassette DraI-DraI 6000bp, MCS-aadA-MCS
FT                   2. pAM28 remove DraI-DraI 19bp, no lacIq gene
FT                   -> plasmid 6000bp
FT                   1. plasmid PstI 6000bp
FT                   fill in
FT                   -> pAM34 6000bp"
FT   -35_signal      55..60
FT                   /note="S35 E. coli lac gene"
FT   -10_signal      79..84
FT                   /note="S10 E. coli lac gene"
FT   promoter        88..114
FT                   /note="PRO E. coli lac operator"
FT   -35_signal      complement(257..262)
FT                   /note="S35 E. coli RNAI gene promoter PI"
FT   -10_signal      complement(281..286)
FT                   /note="S10 E. coli RNAI gene promoter PI"
FT   rep_origin      691..694
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   terminator      complement(1784..1815)
FT                   /note="TER bacteriophage T4 gene 32"
FT   CDS             complement(1120..1311)
FT                   /note="REP E. coli Rom gene (Rop)"
FT   -35_signal      2208..2213
FT                   /note="S35 E. coli aadA gene"
FT   -10_signal      2231..2236
FT                   /note="S10 E. coli aadA gene"
FT   CDS             2253..3263
FT                   /note="ANT E. coli spectinomycin resistance gene
FT                   (spc); aadA gene"
FT   misc_feature    3511..3547
FT                   /note="MCS"
FT   terminator      3596..3627
FT                   /note="TER bacteriophage T4 gene 32"
FT   CDS             complement(3707..4567)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_feature    4023..4023
FT                   /note="base is A, as in pUC plasmids"
FT   -10_signal      complement(4609..4614)
FT                   /note="S10 E. coli bla gene"
FT   -35_signal      4632..4637
FT                   /note="S35 E. coli bla gene"
FT   CDS             complement(4780..5862)
FT                   /note="REP E. coli lacIq repressor gene"
FT   -10_signal      5899..5904
FT                   /note="S10 E. coli lacI gene"
FT   -35_signal      5922..5927
FT                   /note="S35 E. coli lacI gene"
FT   misc_feature    5926..5926
FT                   /note="E. coli lacIq gene mutation"
XX
SQ   Sequence 6000 BP; 1440 A; 1553 C; 1565 G; 1442 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 cacgaggccc tttcgtcttc aagggtcgct cactgcccgc tttccagtcg
     ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     gcgaaaatcc tgtttgatgg tggttgacgg cgggatataa catgagctgt cttcggtatc
     gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc
     ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
//