LOCUS       pTW097_Lvl0_-98/        2877 bp    DNA     circular     27-MAR-2014
DEFINITION  No BsaI or SapI.
ACCESSION   urn.local...hy-9e4hqcb
KEYWORDS    .
SOURCE      Cloning vector pUC19
  ORGANISM  Cloning vector pUC19
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 2686)
  AUTHORS   New, England and Biolabs.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-OCT-2007) Research Department, New England Biolabs,
            240 County Road, Ipswich, MA 01938, USA COMMENT See also GenBank
            accession L09137.
FEATURES             Location/Qualifiers
     source          1..2877
                     /organism="Cloning vector pUC19"
                     /mol_type="other DNA"
     gene            55..655
                     /standard_name="lacZ PCR Product from pUC19"
     primer_bind     55..75
                     /standard_name="TK_DI_lacZ_Fwd_1"
     CDS             complement(146..469)
                     /gene="lacZalpha"
                     /codon_start=1
                     /product="beta-galactosidase alpha fragment"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLN
                     RLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTI
                     CSDAA"
     gene            complement(146..469)
                     /gene="lacZalpha"
     primer_bind     364..386
                     /standard_name="M13/pUC Forward (Quintara)"
     primer_bind     378..395
                     /standard_name="M13 (-21) Forward (Quintara)"
     misc_feature    396..452
                     /gene="lacZalpha"
                     /note="multiple cloning site (EcoRI-HindIII)"
                     /note="Geneious type: polylinker"
     primer_bind     complement(465..481)
                     /standard_name="M13 Reverse (Quintara)"
     -10_signal      complement(514..519)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
     -35_signal      complement(538..543)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
     protein_bind    complement(563..575)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
     primer_bind     578..598
                     /standard_name="TW_lvl0_seq_Fwd1"
     misc_feature    679..693
                     /standard_name="15 bp SapI Overhang"
     misc_feature    694..699
                     /standard_name="BsaI"
     primer_bind     694..881
                     /standard_name="TW_lvl0_98-3_NA_Fwd"
     primer_bind     complement(694..881)
                     /standard_name="TW_lvl0_98-3_NA_Rev"
     misc_feature    701..704
                     /standard_name="BsaI Cut Site"
     misc_feature    702..>802
                     /standard_name="98/5 SGP"
     primer_bind     719..740
                     /standard_name="TK-VEE-nsP4-seq-F"
     primer_bind     755..769
                     /standard_name="ApaI 15 bp Overhang"
     misc_feature    800..>802
                     /standard_name="Subgenomic RNA 5'UTR"
     misc_feature    804..868
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="ACTACGACATAGTCTAGTCCGCCAAG"
     misc_feature    complement(871..874)
                     /standard_name="BsaI Cut Site"
     misc_feature    complement(876..881)
                     /standard_name="BsaI"
     primer_bind     complement(967..987)
                     /standard_name="TW_lvl0_seq_Rev1"
     rep_origin      complement(999..1672)
     misc_RNA        complement(1058..1610)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(1058..1646)
     rep_origin      complement(1058..1591)
     primer_bind     complement(1097..1117)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     1098..1119
                     /standard_name="pHAGE-A7550-7050"
     primer_bind     complement(1362..1386)
                     /standard_name="pHAGE-S7501-8000"
     primer_bind     complement(1406..1425)
                     /standard_name="RepO-R"
     -35_signal      1464..1469
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      1486..1491
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        1500..1607
                     /note="RNAI transcript"
     primer_bind     1609..1630
                     /standard_name="pHAGE-A7050-6550"
     -10_signal      complement(1620..1625)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(1641..1646)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(1817..2677)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(1817..2677)
                     /gene="bla"
     primer_bind     complement(1866..1890)
                     /standard_name="pHAGE-S7001-7500"
     misc_feature    complement(1957..1962)
                     /standard_name="BsaI site"
     primer_bind     2118..2141
                     /standard_name="pHAGE-A6550-6050"
     primer_bind     complement(2386..2410)
                     /standard_name="pHAGE-S6501-7000"
     primer_bind     2440..2459
                     /standard_name="Amp-R; Quintara AmpR"
     primer_bind     2587..2610
                     /standard_name="pHAGE-A6050-5550"
     sig_peptide     complement(2609..2677)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     promoter        complement(2678..2712)
                     /note="Beta Lactamase Promoter"
                     /standard_name="P-Bla"
     -10_signal      complement(2721..2726)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(2742..2747)
                     /note="bla promoter (counter-clockwise); TTCAAA"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
      481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
      661 agaggcggtt tgcgtattgg gcgctcttcc gctggtctcc gacttccatc atagttatgg
      721 ccatgactac tctagctagc agtgttaaat cattcagcta cctgagaggg gcccctataa
      781 ctctctacgg ctaacctgaa tggggcgtga tccgaaaggt gacccggatc tggggcgtga
      841 tccgaaaggt gacccggatc caccggtcgc cacctgagac cgcttcctcg ctcactgact
      901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1741 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1801 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1861 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1921 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1981 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2041 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2101 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2161 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2221 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2281 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2341 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2401 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2461 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2521 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2581 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2641 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     2701 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2761 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     2821 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
