LOCUS       pTW065_Lvl0_3'UT        4662 bp    DNA     circular     10-JAN-2014
DEFINITION  Cloning vector pUC19, complete sequence.
ACCESSION   urn.local...hj-9e4gkg2
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..4662
                     /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"
     primer_bind     659..679
                     /standard_name="TW_lvl0_seq_F"
     misc_feature    679..693
                     /standard_name="15 bp SapI Overhang"
     primer_bind     679..737
                     /standard_name="TW_lvl0_3'UTR_Fwd"
     misc_feature    694..699
                     /standard_name="BsaI"
     misc_feature    701..704
                     /standard_name="BsaI Cut Site"
     misc_feature    705..733
                     /standard_name="Misc repeat 1 in pDONR221"
     misc_feature    732..758
                     /standard_name="Misc repeat 2 in pDONR221"
     misc_feature    753..781
                     /standard_name="Misc repeat 1 in pDONR221"
     gene            complement(1100..1405)
                     /note="Geneious type: Gene"
                     /standard_name="ccdB"
     CDS             complement(1728..2408)
                     /standard_name="Chloramphenicol resistance gene"
     primer_bind     complement(2626..2681)
                     /standard_name="TW_lvl0_3'UTR_Rev"
     misc_feature    2629..2655
                     /standard_name="Misc repeat 2 in pDONR221"
     3'UTR           <2656..2658
     misc_feature    complement(2656..2659)
                     /standard_name="BsaI Cut Site"
     misc_feature    <2656..2658
                     /standard_name="19 nucleotide conserved sequence"
     misc_feature    complement(2661..2666)
                     /standard_name="BsaI"
     primer_bind     2667..2681
                     /standard_name="15 bp SapI Overhang"
     primer_bind     complement(2701..2721)
                     /standard_name="TW_lvl0_seq_R"
     rep_origin      complement(2784..3457)
     misc_RNA        complement(2843..3395)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(2843..3431)
     primer_bind     complement(2882..2902)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     complement(3191..3210)
                     /standard_name="RepO-R"
     -35_signal      3249..3254
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      3271..3276
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        3285..3392
                     /note="RNAI transcript"
     -10_signal      complement(3405..3410)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(3426..3431)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(3602..4462)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(3602..4462)
                     /gene="bla"
     misc_feature    complement(3742..3747)
                     /standard_name="BsaI site"
     primer_bind     4225..4244
                     /standard_name="Amp-R; Quintara AmpR"
     sig_peptide     complement(4394..4462)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     -10_signal      complement(4506..4511)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(4527..4532)
                     /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 ataagtcgac cgacagcctt
      721 ccaaatgttc ttcgggtgat gctgccaact tagtcgaccg acagccttcc aaatgttctt
      781 ctcaaacgga atcgtcgtat ccagcctact cgctattgtc ctcaatgccg tattaaatca
      841 taaaaagaaa taagaaaaag aggtgcgagc ctcttttttg tgtgacaaaa taaaaacatc
      901 tacctattca tatacgctag tgtcatagtc ctgaaaatca tctgcatcaa gaacaatttc
      961 acaactctta tacttttctc ttacaagtcg ttcggcttca tctggatttt cagcctctat
     1021 acttactaaa cgtgataaag tttctgtaat ttctactgta tcgacctgca gactggctgt
     1081 gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
     1141 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
     1201 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
     1261 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
     1321 cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
     1381 tataggtgta aaccttaaac tgcatttcac cagcccctgt tctcgtcagc aaaagagccg
     1441 ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt
     1501 tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg
     1561 acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct
     1621 gcttcatagc atacctcttt ttgacatact tcgggtatac atatcagtat atattcttat
     1681 accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc
     1741 acgcggcgtt tacgcccccc ctgccactca tcgcagtact gttgtaattc attaagcatt
     1801 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     1861 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     1921 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     1981 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     2041 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     2101 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     2161 accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca
     2221 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     2281 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     2341 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     2401 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     2461 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
     2521 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
     2581 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
     2641 gccaacttag tcgacttcat gagaccgctt cctcgctcac tgactcgctg cgctcggtcg
     2701 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2761 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2821 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2881 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3181 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3241 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3301 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3361 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3421 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3481 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3541 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3601 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3661 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3721 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3781 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3841 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3901 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3961 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4021 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4081 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4141 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4201 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4261 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4321 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4381 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4441 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4501 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4561 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4621 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
