LOCUS       pTW095_Lvl0_-98/        2824 bp    DNA     circular     27-MAR-2014
DEFINITION  No BsaI or SapI.
ACCESSION   urn.local...hw-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..2824
                     /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..828
                     /standard_name="TW_lvl0_98-30_NA_Fwd"
     primer_bind     complement(694..828)
                     /standard_name="TW_lvl0_98-30_NA_Rev"
     misc_feature    701..704
                     /standard_name="BsaI Cut Site"
     misc_feature    702..804
                     /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..>815
                     /standard_name="Subgenomic RNA 5'UTR"
     misc_feature    815^816
                     /note="Geneious type: Editing History Deletion"
                     /standard_name="TCTAGTCCGCCAAG"
     misc_feature    complement(818..821)
                     /standard_name="BsaI Cut Site"
     misc_feature    complement(823..828)
                     /standard_name="BsaI"
     primer_bind     complement(914..934)
                     /standard_name="TW_lvl0_seq_Rev1"
     rep_origin      complement(946..1619)
     misc_RNA        complement(1005..1557)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(1005..1593)
     rep_origin      complement(1005..1538)
     primer_bind     complement(1044..1064)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     1045..1066
                     /standard_name="pHAGE-A7550-7050"
     primer_bind     complement(1309..1333)
                     /standard_name="pHAGE-S7501-8000"
     primer_bind     complement(1353..1372)
                     /standard_name="RepO-R"
     -35_signal      1411..1416
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      1433..1438
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        1447..1554
                     /note="RNAI transcript"
     primer_bind     1556..1577
                     /standard_name="pHAGE-A7050-6550"
     -10_signal      complement(1567..1572)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(1588..1593)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(1764..2624)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(1764..2624)
                     /gene="bla"
     primer_bind     complement(1813..1837)
                     /standard_name="pHAGE-S7001-7500"
     misc_feature    complement(1904..1909)
                     /standard_name="BsaI site"
     primer_bind     2065..2088
                     /standard_name="pHAGE-A6550-6050"
     primer_bind     complement(2333..2357)
                     /standard_name="pHAGE-S6501-7000"
     primer_bind     2387..2406
                     /standard_name="Amp-R; Quintara AmpR"
     primer_bind     2534..2557
                     /standard_name="pHAGE-A6050-5550"
     sig_peptide     complement(2556..2624)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     promoter        complement(2625..2659)
                     /note="Beta Lactamase Promoter"
                     /standard_name="P-Bla"
     -10_signal      complement(2668..2673)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(2689..2694)
                     /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 tggactacga cataggccac ctgagaccgc ttcctcgctc
      841 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      901 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      961 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1021 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1081 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1141 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1201 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1261 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1321 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1381 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1441 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1501 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1561 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1621 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1681 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1741 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1801 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1861 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1921 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1981 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2041 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2101 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2161 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2221 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2281 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2341 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2401 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2461 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2521 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2581 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2641 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2701 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     2761 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     2821 cgtc
//
