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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)
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DE E. coli plasmid vector pAM34 - 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]
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
//