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ID   PUR250     preliminary; circular DNA; SYN; 2585 BP.
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AC   ATCC37142;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pUR250 - 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   pUR245 from pUC9 & linker
RC   plasmid from pUR245 & pUR222
RC   pUR250 from pUR219 & plasmid
RA   Ruther U.;
RT   "pUR 250 allows rapid chemical sequencing of both DNA strands of
RT   its inserts";
RL   Nucleic Acids Res. 10:5765-5772(1982).
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CC   Created by Moore, July 1995, under contract with NCBI.
CC   Plasmid vector allowing rapid chemical sequencing of both strands of
CC   inserted DNA. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pUR250)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pUC9)(pUR222)(pUC4)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC9 remove HindIII-PstI-SalI 14bp
FT                   \ 234..248, MCS/2651bp
FT                   2. HindIII-XbaI-SalI linker 12bp agctttctagag
FT                   -> pUR245 2663bp
FT                   1. pUR222 HaeII-BamHI 536bp 1303..1839, lacZ alpha
FT                   \ pUR222 HaeII 1303 1673 2091
FT                   2. pUR245 BamHI-HaeII 250bp 5..-..254, lacPO
FT                   -> fragment 786bp [HaeII-HaeII]
FT                   1. pUC4 HindII 2605bp 2159..2159
FT                   mutagenesis
FT                   remove AccI-AccI 139bp 343..482, 2466bp
FT                   Klenow
FT                   remove HaeII-HaeII 370bp 603..973, 2096bp
FT                   \ pUC4 HaeII = 170 603 973
FT                   -> pUR219 2096bp [no HindII, no AccI, no PstI]
FT                   1. pUR219 remove small HaeII-HaeII 294bp,
FT                   \ pUC4 170..343 & 482..603/1802bp
FT                   2. fragment HaeII-HaeII 786bp, MCS/lacPO/lacZ alpha
FT                   -> pUR250 2588bp"
FT   -               1..169
FT                   /note="pUC4 1..169 169bp
FT                   HaeII = RGCGC^Y"
FT   -               170..705
FT                   /note="pUR222 1303..1838 536bp
FT                   BamHI = G^GATCC"
FT   -               706..934
FT                   /note="pUC9 5..233 229bp
FT                   HindIII = A^AGCTT
FT                   \           agctttctagag"
FT   -               935..946
FT                   /note="agctttctagag 12bp
FT                   \ agctttctagag
FT                   SalI =       G^TCGAC"
FT   -               947..952
FT                   /note="pUC9 248..253 6bp
FT                   HaeII = RGCGC^Y"
FT   -               953..2138
FT                   /note="pUC4 973..2158 1186bp"
FT   -               2139..2585
FT                   /note="pUC4 2159..2605 447bp"
FT   misc_binding    0..0
FT                   /note="MCS HindIII-XbaI-SalI-BamHI-SmaI-EcoRI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 2585 BP; 640 A; 633 C; 667 G; 645 T; 0 other;
     gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc acgcttcccg
     aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     attcgccatt caggctacgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     tacgccagct ggcgaagggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ccccgccaat acgcaaaccg
     cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     cacacaggaa acagctatga ccatgattac gccaagcttt ctagagtcga cgtttctcaa
     tgctcacgct 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 ctacgggata 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
     tcgtc
//