LOCUS       pTW120_Lvl0_TetR        3334 bp    DNA     circular     27-MAR-2014
DEFINITION  1 BsaI and SapI Sites.
ACCESSION   urn.local...k6-9e4hqe1
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..3334
                     /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     complement(637..655)
                     /standard_name="TK_DI_lacZ_Rev_1"
     misc_feature    679..693
                     /standard_name="15 bp SapI Overhang"
     primer_bind     679..>704
                     /standard_name="TW_lvl0_ORF_Fwd"
     misc_feature    694..699
                     /standard_name="BsaI"
     primer_bind     694..722
                     /standard_name="TW_lvl0_TetR_Fwd"
     misc_feature    701..704
                     /standard_name="BsaI Cut Site"
     gene            705..1328
                     /standard_name="TetR"
     primer_bind     complement(1307..1338)
                     /standard_name="TW_lvl0_TetR_Rev"
     gene            1328
                     /standard_name="mVenus"
     misc_feature    complement(1328..1331)
                     /standard_name="BsaI Cut Site"
     primer_bind     complement(<1328..1353)
                     /standard_name="TW_lvl0_ORF_Rev"
     misc_feature    complement(1333..1338)
                     /standard_name="BsaI"
     primer_bind     1339..1353
                     /standard_name="15 bp SapI Overhang"
     rep_origin      complement(1456..2129)
     misc_RNA        complement(1515..2067)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(1515..2103)
     primer_bind     complement(1554..1574)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     complement(1863..1882)
                     /standard_name="RepO-R"
     -35_signal      1921..1926
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      1943..1948
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        1957..2064
                     /note="RNAI transcript"
     -10_signal      complement(2077..2082)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(2098..2103)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(2274..3134)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(2274..3134)
                     /gene="bla"
     misc_feature    complement(2414..2419)
                     /standard_name="BsaI site"
     primer_bind     2897..2916
                     /standard_name="Amp-R; Quintara AmpR"
     sig_peptide     complement(3066..3134)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     -10_signal      complement(3178..3183)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(3199..3204)
                     /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 gctggtctca caccatgtca agactcgaca
      721 agagcaaggt gattaacagt gcactggaac ttctcaatga agttgggatc gaggggctga
      781 ctactagaaa actcgcacag aaactggggg ttgagcagcc caccttgtac tggcacgtta
      841 aaaacaaaag ggccctgctg gatgctctgg ccatcgagat gctggatagg catcataccc
      901 acttctgccc tctggaagga gaatcctggc aggatttcct tagaaacaac gccaagtcct
      961 ttcgctgtgc tcttcttagc caccgggatg gtgctaaagt ccatctcggc acacgaccaa
     1021 ctgagaagca gtacgaaact ctcgagaacc agctggcctt tctctgtcaa cagggctttt
     1081 ctcttgaaaa cgccctgtac gcactgagtg cagttgggca ctttacactc ggatgtgttc
     1141 tggaggacca agaacatcag gtggcaaagg aagagaggga gacccctacg actgactcca
     1201 tgccccctct cttgaggcag gcaatagaat tgttcgacca tcagggcgca gaacccgcct
     1261 ttctgtttgg gctggaactg attatctgcg gtcttgagaa acagctgaag tgcgagtccg
     1321 ggagctaata atgagaccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1381 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1441 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1501 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1561 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1621 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1681 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1741 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1801 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1861 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1921 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1981 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2041 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2101 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2161 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2221 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2281 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2341 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2401 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2461 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2521 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2581 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2641 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2701 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2761 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2821 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2881 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2941 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3001 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3061 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3121 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3181 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3241 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3301 tataaaaata ggcgtatcac gaggcccttt cgtc
//
