LOCUS       pTW094_Lvl0_E1-3        3038 bp    DNA     circular     27-MAR-2014
DEFINITION  No BsaI or SapI.
ACCESSION   urn.local...it-9e4hqcx
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..3038
                     /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"
     misc_feature    694..699
                     /standard_name="BsaI"
     primer_bind     694..1042
                     /standard_name="TW_lvl0_3UTR_Fwd"
     primer_bind     complement(694..1042)
                     /standard_name="TW_lvl0_3UTR_Rev"
     misc_feature    701..704
                     /standard_name="BsaI Cut Site"
     gene            705..914
                     /standard_name="E1 (truncated)"
     primer_bind     complement(775..794)
                     /standard_name="TK-VEE-E1-seq-R"
     3'UTR           918..1034
     primer_bind     complement(959..978)
                     /standard_name="TW_VEE_Seq_1_R"
     primer_bind     complement(965..987)
                     /standard_name="TK-VEE-3'UTR-seq-R"
     misc_feature    1016..1034
                     /standard_name="19 nucleotide conserved sequence"
     misc_feature    complement(1032..1035)
                     /standard_name="BsaI Cut Site"
     misc_feature    complement(1037..1042)
                     /standard_name="BsaI"
     primer_bind     1043..1057
                     /standard_name="15 bp SapI Overhang"
     rep_origin      complement(1160..1833)
     misc_RNA        complement(1219..1771)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(1219..1807)
     rep_origin      complement(1219..1752)
     primer_bind     complement(1258..1278)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     1259..1280
                     /standard_name="pHAGE-A7550-7050"
     primer_bind     complement(1523..1547)
                     /standard_name="pHAGE-S7501-8000"
     primer_bind     complement(1567..1586)
                     /standard_name="RepO-R"
     -35_signal      1625..1630
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      1647..1652
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        1661..1768
                     /note="RNAI transcript"
     primer_bind     1770..1791
                     /standard_name="pHAGE-A7050-6550"
     -10_signal      complement(1781..1786)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(1802..1807)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(1978..2838)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(1978..2838)
                     /gene="bla"
     primer_bind     complement(2027..2051)
                     /standard_name="pHAGE-S7001-7500"
     misc_feature    complement(2118..2123)
                     /standard_name="BsaI site"
     primer_bind     2279..2302
                     /standard_name="pHAGE-A6550-6050"
     primer_bind     complement(2547..2571)
                     /standard_name="pHAGE-S6501-7000"
     primer_bind     2601..2620
                     /standard_name="Amp-R; Quintara AmpR"
     primer_bind     2748..2771
                     /standard_name="pHAGE-A6050-5550"
     sig_peptide     complement(2770..2838)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     promoter        complement(2839..2873)
                     /note="Beta Lactamase Promoter"
                     /standard_name="P-Bla"
     -10_signal      complement(2882..2887)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(2903..2908)
                     /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 ataatatgtt acgtgcaaag
      721 gtgattgtca ccccccgaaa gaccatattg tgacacaccc tcagtatcac gcccaaacat
      781 ttacagccgc ggtgtcaaaa accgcgtgga cgtggttaac atccctgctg ggaggatcag
      841 ccgtaattat tataattggc ttggtgctgg ctactattgt ggccatgtac gtgctgacca
      901 accagaaaca taattgaata cagcagcaat tggcaagctg cttacataga actcgcggcg
      961 attggcatgc cgccttaaaa tttttatttt atttttcttt tcttttccga atcggatttt
     1021 gtttttaata tttcatgaga ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1621 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2581 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2821 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2941 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3001 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
