LOCUS       pTW098_Lvl0_-98/        2879 bp    DNA     circular     27-MAR-2014
DEFINITION  No BsaI or SapI.
ACCESSION   urn.local...hz-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..2879
                     /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..883
                     /standard_name="TW_lvl0_98-5_NA_Fwd"
     primer_bind     complement(694..883)
                     /standard_name="TW_lvl0_98-5_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..>804
                     /standard_name="Subgenomic RNA 5'UTR"
     misc_feature    805..870
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="CTACGACATAGTCTAGTCCGCCAAG"
     misc_feature    complement(873..876)
                     /standard_name="BsaI Cut Site"
     misc_feature    complement(878..883)
                     /standard_name="BsaI"
     primer_bind     complement(969..989)
                     /standard_name="TW_lvl0_seq_Rev1"
     rep_origin      complement(1001..1674)
     misc_RNA        complement(1060..1612)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(1060..1648)
     rep_origin      complement(1060..1593)
     primer_bind     complement(1099..1119)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     1100..1121
                     /standard_name="pHAGE-A7550-7050"
     primer_bind     complement(1364..1388)
                     /standard_name="pHAGE-S7501-8000"
     primer_bind     complement(1408..1427)
                     /standard_name="RepO-R"
     -35_signal      1466..1471
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      1488..1493
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        1502..1609
                     /note="RNAI transcript"
     primer_bind     1611..1632
                     /standard_name="pHAGE-A7050-6550"
     -10_signal      complement(1622..1627)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(1643..1648)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(1819..2679)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(1819..2679)
                     /gene="bla"
     primer_bind     complement(1868..1892)
                     /standard_name="pHAGE-S7001-7500"
     misc_feature    complement(1959..1964)
                     /standard_name="BsaI site"
     primer_bind     2120..2143
                     /standard_name="pHAGE-A6550-6050"
     primer_bind     complement(2388..2412)
                     /standard_name="pHAGE-S6501-7000"
     primer_bind     2442..2461
                     /standard_name="Amp-R; Quintara AmpR"
     primer_bind     2589..2612
                     /standard_name="pHAGE-A6050-5550"
     sig_peptide     complement(2611..2679)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     promoter        complement(2680..2714)
                     /note="Beta Lactamase Promoter"
                     /standard_name="P-Bla"
     -10_signal      complement(2723..2728)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(2744..2749)
                     /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 tggagggcgt gatccgaaag gtgacccgga tctggggcgt
      841 gatccgaaag gtgacccgga tccaccggtc gccacctgag accgcttcct cgctcactga
      901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1921 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2101 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2401 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2821 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
