LOCUS       pTW101_Lvl0_-98/        2890 bp    DNA     circular     27-MAR-2014
DEFINITION  No BsaI or SapI.
ACCESSION   urn.local...i2-9e4hqcc
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..2890
                     /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..894
                     /standard_name="TW_lvl0_98-30_NA_Fwd"
     primer_bind     complement(694..894)
                     /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    816..881
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="TCTAGTCCGCCAAG"
     misc_feature    complement(884..887)
                     /standard_name="BsaI Cut Site"
     misc_feature    complement(889..894)
                     /standard_name="BsaI"
     primer_bind     complement(980..1000)
                     /standard_name="TW_lvl0_seq_Rev1"
     rep_origin      complement(1012..1685)
     misc_RNA        complement(1071..1623)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(1071..1659)
     rep_origin      complement(1071..1604)
     primer_bind     complement(1110..1130)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     1111..1132
                     /standard_name="pHAGE-A7550-7050"
     primer_bind     complement(1375..1399)
                     /standard_name="pHAGE-S7501-8000"
     primer_bind     complement(1419..1438)
                     /standard_name="RepO-R"
     -35_signal      1477..1482
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      1499..1504
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        1513..1620
                     /note="RNAI transcript"
     primer_bind     1622..1643
                     /standard_name="pHAGE-A7050-6550"
     -10_signal      complement(1633..1638)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(1654..1659)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(1830..2690)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(1830..2690)
                     /gene="bla"
     primer_bind     complement(1879..1903)
                     /standard_name="pHAGE-S7001-7500"
     misc_feature    complement(1970..1975)
                     /standard_name="BsaI site"
     primer_bind     2131..2154
                     /standard_name="pHAGE-A6550-6050"
     primer_bind     complement(2399..2423)
                     /standard_name="pHAGE-S6501-7000"
     primer_bind     2453..2472
                     /standard_name="Amp-R; Quintara AmpR"
     primer_bind     2600..2623
                     /standard_name="pHAGE-A6050-5550"
     sig_peptide     complement(2622..2690)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     promoter        complement(2691..2725)
                     /note="Beta Lactamase Promoter"
                     /standard_name="P-Bla"
     -10_signal      complement(2734..2739)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(2755..2760)
                     /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 cataggggcg tgatccgaaa ggtgacccgg
      841 atctggggcg tgatccgaaa ggtgacccgg atccaccggt cgccacctga gaccgcttcc
      901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1501 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2821 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2881 ccctttcgtc
//
