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ID   PSP65      preliminary; circular DNA; SYN; 3005 BP.
XX
AC   X65329; L08944; VB0035; IG0096;
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DT   06-AUG-1987 (Rel. 2, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli phagemid vector pSP65 - 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   pSP65
RA   ;
RT   ;
RL   Submitted (23-MAR-1992) on tape to the EMBL Data Library by:
RL   Technical Services, Promega Corporation, 2800 Woods Hollow Road,
RL   Madison, WI 53711-5399, USA.
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RN   [2]
RP   1-3005
RC   pSP65
RA   Klock C.;
RT   ;
RL   Submitted (28-MAY-1993) on tape to the EMBL Data Library by:
RL   Technical Services, Promega Corporation, 2800 Woods Hollow Road,
RL   Madison, WI 53711-5399, USA.
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RN   [3]
RC   pSP64 from pUC12 & SP6 promoter
RC   pSP65 from pUC13 & SP6 promoter
RA   Melton D.A., Krieg P.A., Rebagliati M.R., Maniatis T., Zinn K.,
RA   Green M.R.;
RT   "Efficient in vitro synthesis of biologically active RNA and RNA
RT   hybridization probes from plasmids containing a bacteriophage
RT   SP6 promoter";
RL   Nucleic Acids Res. 12:7035-7056(1984).
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RN   [4]
RC   pSP65 from pUC13 & SP6 promoter
RA   Melton D.A.;
RT   ;
RL   Unpublished (1986).
RL   Melton D.A., Harvard University.
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RN   [5]
RC   pSP65
RA   Gilbert W.;
RT   "Obtained from VecBase 3.0";
RL   Unpublished (1991).
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CC   During the construction of the pSP65 vector from pUC12, the
CC   majority of the lacZ gene was deleted.  Therefore. the X-gal
CC   color screening method for the identification of recombinants is
CC   not possible with this vector.
CC   old Boehringer vector.
CC   NM (pSP65)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (Promega)
CC   HO (E.coli HB101)(E.coli LE392)(E.coli JM83)
CC   CP (high)
CC   FN (transcription)
CC   SE (color blue/white)
CC   PA (pUC13)(bGal13)(PromSP6)(pBR322)
CC   BR (pSP64)
CC   OF (pGEM-2)
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC13 remove small FnuDII-Sau3A/MboI,
FT                   \ 2680bp-
FT                   2. SP6 FnuDII-Sau3A/MboI 400bp, promoter
FT                   -> pSP65 3005bp [no lacZ]"
FT   promoter        1..1
FT                   /note="PRO Salmonella bacteriophage Sp6
FT                   transcription initiation site"
FT   misc_binding    10..60
FT                   /note="MCS EcoRI-SacI-SmaI-BamHI-XbaI-SalI-PstI-
FT                   HindIII, from M13mp11/pUC13"
FT   misc_feature    63..292
FT                   /note="E. coli lac operon"
FT   misc_feature    64..2294
FT                   /note="from pUC13 bases 450..2680"
FT   promoter        97..113
FT                   /note="PRO E. coli lac operator"
FT   misc_feature    77..77
FT                   /note="E. coli lacZ gene transcription initiation
FT                   site"
FT   misc_binding    238..238
FT                   /note="SIT unique PvuII"
FT   misc_binding    414..414
FT                   /note="SIT unique AflIII"
FT   rep_origin      <414..<1000
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             1234..2094
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_binding    1427..1427
FT                   /note="SIT unique BglI"
FT   misc_binding    1529..1529
FT                   /note="SIT unique AvlII"
FT   misc_binding    1677..1677
FT                   /note="SIT unique PvuII"
FT   misc_binding    1787..1787
FT                   /note="SIT unique ScaI"
FT   misc_binding    1906..1906
FT                   /note="SIT unique Asp700I"
FT   misc_binding    2111..2111
FT                   /note="SIT unique SspI"
FT   misc_binding    2229..2229
FT                   /note="SIT unique AatII"
FT   misc_feature    2295..2481
FT                   /note="from pUC13 bases 1..187"
FT   misc_feature    2528..2806
FT                   /note="from pBR322 bases complement(376..654)"
FT   misc_binding    2574..2574
FT                   /note="SIT unique SphI"
FT   misc_binding    2603..2603
FT                   /note="SIT unique DsaI"
FT   misc_binding    2639..2639
FT                   /note="SIT unique Eco47III"
FT   misc_binding    2732..2732
FT                   /note="SIT unique NaeI"
FT   misc_binding    2763..2763
FT                   /note="SIT unique NheI"
FT   misc_feature    2807..3005
FT                   /note="from bacteriophage Sp6"
FT   promoter        join(2989..3005,1..6)
FT                   /note="PRO bacteriophage Sp6; from Salmonella"
XX
SQ   Sequence 3005 BP; 745 A; 777 C; 761 G; 722 T; 0 other;
     gaatacacgg aattcgagct cgcccgggga tcctctagag tcgacctgca gcccaagctt
     ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     tgtactgaga gtgcaccata tcgacgctct cccttatgcg actcctgcat taggaagcag
     cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     tggctagcga tgaccctgct gattggttcg ctgaccattt ccggggtgcg gaacggcgtt
     accagaaact cagaaggttc gtccaaccaa accgactctg acggcagttt acgagagaga
     tgatagggtc tgcttcagta agccagatgc tacacaatta ggcttgtaca tattgtcgtt
     agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
     ctata
//