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ID   PTRS3      preliminary; circular DNA; SYN; 3930 BP.
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AC   L08955; K01480; VB0086;
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DT   07-JUN-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pTrS3 - complete.
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KW   cloning vector; promoter; trp promoter; SD sequence; ATG.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-3930
RC   pTrS3
RA   Gilbert W.;
RT   "Obtained from VecBase 3.0";
RL   Unpublished (1991).
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RN   [2]
RP   1-3930
RC   [lambda cI857 trpED10 from lambda, cI857 gene/trpED10 gene]
RC   pKYP1 from lambda cI857 trpED10 & pBR325
RC   pKYP5 from pKYP1 & pBR322
RC   pKYP10 from pKYP5 & pBR322
RC   pKYP100 from pKYP10 & pBR322
RC   pTrS3 from pKYP100 & oligo
RC   pLE3 from pBR322 & human beta-interferon gene
RC   pFJ117, pFJ118, pFJ123 from pTrS3 & pLE3
RC   pFM9 from pTrS3 & pLE3
RA   Nishi T., Sato M., Saito A., Itoh S., Takaoka C., Taniguchi T.;
RT   "Construction and application of a novel plasmid ATG vector
RT   for direct expression of foreign genes in Escherichia coli";
RL   DNA 2:265-273(1983).
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RN   [3]
RC   from lambda cI857 trpED10
RC   pGM1 from pVH153
RC   pGM3 from pVH153
RA   Miozzari G.F., Yanofsky C.;
RT   "Tranlation of the leader region of the Escherichia coli
RT   tryptophan operon";
RL   J. Bacteriol. 133:1457-1466(1978).
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RN   [4]
RC   pVH153 from pVH51 & lambda cI857 trpED10
RA   Hershfield V., Helinski D.;
RT   ;
RL   Unpublished (1978).
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RN   [5]
RC   lambda cI857 trpED10 from lambda,  trpED10 gene/cI857 gene
RA   Helinski D.R., Hershfield V., Figurski D., Meyer R.J.;
RT   "Construction and properties of plasmid cloning vehicles";
RL   Recombinant Molecules 0:151-165(1977).
RL   Beers R.F., Basset E.G., Raven Press, New York.
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CC   These data and their annotation were supplied to GenBank by Will
CC   Gilbert under the auspices of the GenBank Currator Program.
CC
CC   Expression of foreign genes in E.coli requires an E. coli promoter
CC   to direct initiation of transcription and a translation initiation
CC   signal that consists of an ATG initiation codon preceeded by a
CC   Shine Dalgarno (SD) sequence. These sequences were created in a
CC   plasmid vector carrying the E.coli tryptophan promoter. The ATG
CC   sequence is 13 bp downstream from the SD sequence. After cleavage
CC   with SphI the 3' ends are blunted using the Klenow fragment of
CC   E. coli DNA polymerase I. Then foreign genes can be ligated to this
CC   vector DNA and cloned in E. coli successfully.
CC   [1] presents successful expression of human interferon-beta in E.coli.
CC   NCBI gi: 310818 & 209195
CC   NM (pTrS3)
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)
CC   SE ()
CC   PA (pKYP100 from pBR322 and lambda-cI857trpED10)
CC   BR ()
CC   OF (pFJ117)(pFJ118)(pFJ123)(pFM9)
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. lambda-cI857trpED10 EcoRI-HindIII 4600bp,trp
FT                   2. pBR322 remove EcoRI-HindIII 31bp 4360..4361..30,
FT                   \ 4330bp
FT                   -> pKYP1 8900bp
FT                   1. pKYP1 TaqI-TaqI, trp prom/SD/no ATG
FT                   EcoRI-TaqI 2600bp
FT                   2. pBR322 remove EcoRI-TaqI 26bp 4360..4361..25,
FT                   \ 4335bp
FT                   -> pKYP5 6900bp
FT                   1. pKYP5 HpaII-HindIII 340bp, trp promoter
FT                   2. pBR322 remove ClaI-HindIII 5bp 25..30, 4356bp
FT                   -> pKYP10 4700bp
FT                   1. pKYP10 HhaI-HhaI 340bp, trp promoter
FT                   blunt end:blunt end
FT                   HindIII-HindIII 109bp, trp promoter
FT                   XhoI linker 6bp ctcgag:XhoI linker 6bp ctcgag
FT                   2. pBR322 HindIII-EcoRI 4330bp 30..4360
FT                   fill in
FT                   -> pKYP100 4439bp
FT                   1. pKYP100 ClaI-SphI 3897bp 567..4439..25
FT                   2. oligo 16bp cgataagctatgcatg
FT                   -> pTrS3 3930bp"
FT   misc_binding    1..6
FT                   /note="SIT EcoRI, from pBR322"
FT   misc_binding    6..21
FT                   /note="SIT XhoI, linker duplet"
FT   promoter        21..123
FT                   /note="PRO E. coli trp"
FT   misc_binding    121..136
FT                   /note="MCS ClaI-ATG-SphI; linker"
FT   misc_binding    0..0
FT                   /note="SIT SalI"
FT   misc_feature    131..3930
FT                   /note="from pBR322"
FT   misc_binding    0..0
FT                   /note="SIT PvuII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             2868..3656
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_binding    0..0
FT                   /note="SIT PstI"
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SQ   Sequence 3930 BP; 929 A; 1066 C; 1005 G; 930 T; 0 other;
     gaattcctcg aggcctcgag gccgacatca taacggttct ggcaaatatt ctgaaatgag
     ctgttgacaa ttaatcatcg aactagttaa ctagtacgca agttcacgta aaaagggtat
     cgataagcta tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat gcccttgaga
     gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag
     cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc gctgatcgtc
     acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc
     gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
     acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga attggagcca
     atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg
     cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg tcctggccac
     gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     tcattacccc catgaacaga aattccccct tacacggagg catcaagtga ccaaacagga
     aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     aggcgtatca cgaggccctt tcgtcttcaa
//