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ID   PNP6COLCT  preliminary; circular DNA; SYN; 8071 BP.
XX
AC   M88537;
XX
DT   25-SEP-1992 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   E. coli plasmid vector pNP6Col-CTAP - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-8071
RC   pNP6dRI from pNP6
RC   pNP6CTAP, pNP6CRM-CTAP, pNP6COL-CTAP, pBR-CRM-CTAP from pNP6dRI
RA   Waleh N.S., Sohel I., Lazar J.B., Hudson D.V., Sze P., Underhill P.,
RA   Johnson P.H.;
RT   "Expression of synthetic genes encoding fused proteins under tight
RT   control of modified regulatory regions of the colicin operon";
RL   Gene 117:7-14(1992).
XX
RN   [2]
RC   pNP4, pNP6, pNP12, pNP14, pNP series from pBR322 & ColE1
RA   Waleh N.S., Johnson P.H.;
RT   "Structural and functional organization of the colicin E1 operon";
RL   Proc. Natl. Acad. Sci. U.S.A. 82:8389-8393(1985).
XX
CC   NM (pNP6Col-CTAP)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (cloning)(expression)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR322
FT                   dC tailing
FT                   2. ColE1 2400bp, cea gene/no Col imm
FT                   dG tailing
FT                   -> pNP6
FT                   1. pNP6 remove EcoRI-EcoRI
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pNP6dRI 7791bp [unique EcoRI 7640..7640, cea gene]
FT                   1. pNP6dRI EcoRI 7640..7640
FT                   2. CTAP-III(Leu2) EcoRI-EcoRI 280bp
FT                   -> pNP6Col-CTAP 8071bp"
FT   -35_signal      6024..6029
FT                   /note="S35 E. coli colicin E1 gene"
FT   -10_signal      6047..6052
FT                   /note="S10 E. coli colicin E1 gene"
FT   promoter        6059..6091
FT                   /note="PRO E. coli colicin E1 gene"
FT   RBS             6123..6127
FT                   /note="RBS E. coli colicin E1 gene"
FT   CDS             6135..7652
FT                   /note="GEN E. coli colicin E1 gene; 506 amino acids,
FT                   partial"
FT   CDS             6135..7910
FT                   /note="GEN E. coli colicin E1 and connective tissue
FT                   activating peptide III small fusion protein"
FT   misc_recomb     7652..7653
FT                   /note="junction between colicin E1/CTAP-III (Leu21)"
FT   mat_peptide     7653..7907
FT                   /note="E. coli connective tissue activating peptide
FT                   III (Leu21 or CTAP-III); putative"
FT   terminator      8006..8037
FT                   /note="TER E. coli Rho-dependent terminator; putative"
XX
SQ   Sequence 8071 BP; 2017 A; 1973 C; 2072 G; 2009 T; 0 other;
     ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     cgtatcacga ggccctttcg tcttcaagaa ttaattctca tgtttgacag cttatcatcg
     ataagcttta atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta
     tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca
     taggcttggt tatgccggta ctgccgggcc tcttgcggga tatcgtccat tccgacagca
     tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac
     ccgttctcgg agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac
     ttggagccac tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atcctctacg
     ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     tgcaggagtc gcataaggga gagcgtcgac cgatgccctt gagagccttc aacccagtca
     gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct
     ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc
     tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta
     tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct
     ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc
     aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
     cggctcttac cagcctaact tcgatcactg gaccgctgat cgtcacggcg atttatgccg
     cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
     gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
     gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
     aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
     gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     cagaaattcc cccttacacg gaggcatcaa gtgaccaaac aggaaaaaac cgcccttaac
     atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
     gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
     agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggg gggggggggg
     gagggggggg ggggggctgg aaacacattt gctgaacgcc ttagagcagc tgcaacagga
     ctatatgcag cggctgagcg aatgggagag cgccttcgtg gaattgcaga agatgttttc
     gcttacgcaa cgggacaacg cgatgctgaa cgagcgggtc atgcagttga gtcagcaggt
     gcagcacttg agcgagcaga cagaacgctt gagccagtta tacagcgaga actggagata
     agagaggaac ggctgataca ggagcgcgaa catgtgttat ccctggaacg ggagcgtcag
     ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa
     tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta
     ttcatacggt taaaatttat caggcgcgat cgcgcagttt ttcgggtggt ttgttgccat
     ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg
     gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagaaatc
     gtgtcagcca gcagcctgga ttttttcggg gtaaggtctt tgctgcggta atcagtaccc
     gcaaattgag ttgtgccccg gagtgagttt aaaaattcct ggctgtgaaa ctgctgaatg
     cgttgttgca catcactccg tgattttcct gtcgctttat ctttagctcc catatcccag
     aactgccgaa aaaaaggcac catcattcgt gttgtgtcgg ctttttgggt ggaaaagttg
     ggatcaaaac cgtcatcaga agcatatacg taatgccggg tatgttgttc cacgctgccc
     tttggggtac aggctgtgag agctactgtt gctgttatca tggcgataag acaaagtttg
     tttccggtac gtctcatgag tgtctcccta ccttaaagta tttattctcg gtgagttatt
     atccgaagac tattttatca tcagtctctg acagcgttca gcgaccgaac aacctggacc
     agaatccccg tttcttttcc agttccagtg cctgcctgag ctgggctatc tcattttgta
     gctgttcccg ttctgcttcc tggcgtctgc gatccatatc ctgtgcctgt ttatcctcaa
     gcatcagcgt caggcattgt ttcagctcat tcagttcccg gagaatgcgt tctgtctgct
     gatcatgtgg attttctgca tgtccctcac tgtgcctttc tggtgtctca ttctgcttta
     attcgccgta tgcccggatc agttcactgg tttcaaactc ccgtcgtccg tccctgccgg
     tacggtaact gacaaggcct gcgcgcatat cacggtaaaa ctgtgaacgc gatctgcctg
     tcatttttag tgcgtcccgg agtgtgtgcc atgccataaa gtgacagtgt cccatagatg
     tctcatctca tagtttcagt aaaacataat gaggtctgag aacggtaatg tttgtgctgg
     tttttgtggc atcgggcgag aatagcgcgt ggtgtgaaag actgtttttt tgatcgtttt
     cacaaaaatg gaagtccaca gtcttgacag ggaaaatgca gcggcgtagc ttttatgctg
     tatataaaac cagtggttat atgtacagta tttattttta acttattgtt ttaaaagtca
     aagaggattt tataatggaa accgcggtag cgtactataa agatggtgtt ccttatgatg
     ataagggaca ggtaattatt actcttttga atggtactcc tgacgggagt ggctctggcg
     gcggaggtgg aaaaggaggc agtaaaagtg aaagttctgc agctattcat gcaactgcta
     aatggtctac tgctcaatta aagaaaacac aggcagagca ggctgcccgg gcaaaagctg
     cagcggaagc acaggcgaaa gcaaaggcaa acagggatgc gctgactcag cgcctgaagg
     atatcgtgaa tgaggctctt cgtcacaatg cctcacgtac gccttcagca acagagcttg
     ctcatgctaa taatgcagct atgcaggcgg aagacgagcg tttgcgcctt gcgaaagcag
     aagaaaaagc ccgtaaagaa gcggaagcag cagaaaaggc ttttcaggaa gcagaacaac
     gacgtaaaga gattgaacgg gagaaggctg aaacagaacg ccagttgaaa ctggctgaag
     ctgaagagaa acgactggct gcattgagtg aagaagctaa agctgttgag atcgcccaaa
     aaaaactttc tgctgcacaa tctgaagtgg tgaaaatgga tggagagatt aagactctca
     attctcgttt aagctccagt atccatgccc gtgatgcaga aatgaaaacg ctcgctggaa
     aacgaaatga actggctcag gcatccgcta aatataaaga actggatgag ctggtcaaaa
     aactatcacc aagagccaat gatccgcttc agaaccgtcc tttttttgaa gcaaccagac
     gacgggttgg ggccggtaag attagagaag aaaaacaaaa acaggtaaca gcatcagaaa
     cacgtattaa ccggataaat gctgatataa ctcagatcca gaaggctatt tctcaggtca
     gtaataatcg taatgccggt atcgctcgtg ttcatgaagc tgaagaaaat ttgaaaaaag
     cacagaataa tctccttaat tcacagatta aggatgctgt tgatgcaaca gttagctttt
     atcaaacgct gactgaaaaa tatggtgaaa aatattcgaa aatggcacag gaacttgctg
     ataagtctaa aggtaagaaa atcggcaatg tgaatgaagc tctcgctgct tttgaaaaat
     acaaggatgt tttaaataag aaattcagca aagccgatcg tgatgctatt tttaatgcgt
     tggcatcggt gaagtatgat gactgggcta aacatttaga tcagtttgcc aagtacttga
     agattacggg gcatgtttct tttggatatg atgtggtatc tgatatccta aaaattaagg
     atacaggtga ctggaagcca ctatttctta cattagagaa gaaagctgca gatgcagggg
     tgagttatgt tgttgcttta ctttttagct tgcttgctgg aactacatta ggtatttggg
     gtattgctat tgttacagga attcagctga tgaacctggc taaaggtaaa gaagaatctc
     tggactctga cttatacgct gaactgcgtt gcctgtgcat caaaactact tctgggatcc
     acccgaaaaa catccagtct ctggaagtta tcggtaaagg cactcactgc aaccaggttg
     aagttatcgc tactctgaaa gacggtcgta aaatctgtct agatccggac gctccacgta
     tcaagaagat cgttcagaaa aaactggctg gtgacgaatc tgctgactaa tgactgcaga
     attctatgct cctatattga taagaataaa cttaatacta taaatgaggt gttagggatt
     taattattct ttattgatat aaaaagtcct agcaatccaa atgggattgc taggaccaaa
     cccccccccc cccccccccc ccccccctgc a
//