LOCUS       V044752                 7767 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044752
VERSION     V044752
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2447
                     /label="KRT6B(NM_005555)"
                     /note="KRT6B(NM_005555)"
                     /gene="KRT6B"
     CDS             2454..2519
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2543..3131
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3197..3245
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3447..3875
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3889..4218
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4285..5079
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5255..5376
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5425..5441)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5449..5465
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5473..5503)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5518..5539
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5827..6415)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6586..7446)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7447..7551)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc cagcacatcc accaccatca
      781 ggagccacag cagcagccgc cggggtttca gtgccaactc agccaggctc cctggggtca
      841 gccgctctgg cttcagcagc atctccgtgt cccgctccag gggcagtggt ggcctgggtg
      901 gcgcatgtgg aggagctggc tttggcagcc gcagtctgta tggcctgggg ggctccaaga
      961 ggatctccat tggagggggc agctgtgcca tcagtggcgg ctatggcagc agagccggag
     1021 gcagctatgg ctttggtggc gccgggagtg gatttggttt cggtggtgga gccggcattg
     1081 gctttggtct gggtggtgga gccggccttg ctggtggctt tgggggccct ggcttccctg
     1141 tgtgcccccc tggaggcatc caagaggtca ctgtcaacca gagtctcctg actcccctca
     1201 acctgcaaat tgaccccgcc atccagcggg tgcgggccga ggagcgtgag cagatcaaga
     1261 ccctcaacaa caagtttgcc tccttcatcg acaaggtgcg gttcctagag cagcagaaca
     1321 aggttctgga caccaagtgg accctgctgc aggagcaggg caccaagact gtgaggcaga
     1381 acctggagcc gttgttcgag cagtacatca acaacctcag gaggcagctg gacaacatcg
     1441 tgggggaacg gggtcgtctg gactcggagc tgagaaacat gcaggacctg gtggaggacc
     1501 tcaagaacaa atatgaggat gaaatcaaca agcgcacagc agcagagaat gaatttgtga
     1561 ctctgaagaa ggatgtggat gctgcctaca tgaacaaggt tgaactgcaa gccaaggcag
     1621 acactcttac agatgagatc aacttcctga gagccttgta tgatgcagag ctgtcccaga
     1681 tgcagaccca catctcagac acatccgtgg tgctatccat ggacaacaac cgcaacctgg
     1741 acctggacag catcatcgct gaggtcaagg cccaatatga ggagattgct cagaggagca
     1801 gggctgaggc tgagtcctgg taccagacaa agtacgagga gctgcagatc acagcaggca
     1861 gacatgggga cgacctgcgc aacaccaagc aggagattgc tgagatcaac cgcatgatcc
     1921 agaggctgag atctgagatc gaccacgtca agaagcagtg tgccaaccta caggccgcca
     1981 ttgctgatgc tgagcagcgt ggggagatgg ccctcaagga tgctaagaac aagctggaag
     2041 ggctggagga tgccctgcag aaggccaagc aggacctggc ccggctgctg aaggagtacc
     2101 aggagctgat gaacgtcaag ctggccctgg atgtggagat cgccacctac cgcaagctgc
     2161 tggagggcga ggagtgcagg ctgaatggcg aaggcgttgg acaagtcaac atctctgtag
     2221 tgcagtccac cgtctccagt ggctatggcg gtgccagcgg tgtcggcagt ggcttaggcc
     2281 tgggtggagg aagcagctac tcctatggca gtggtcttgg cgttggaggc ggctttagtt
     2341 ccagcagcgg cagagccact gggggtggcc tcagctctgt tggaggcggc agttccacca
     2401 tcaagtacac caccacctcc tcctccagca ggaagagcta caagcacgga tccgactaca
     2461 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2521 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2581 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2641 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2701 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2761 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2821 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2881 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2941 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3001 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3061 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3121 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3181 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3241 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3301 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3361 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3421 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3481 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3541 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3601 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3661 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3721 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3781 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3841 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3901 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3961 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4021 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4081 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4141 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4201 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4261 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4321 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4381 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4441 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4501 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4561 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4621 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4681 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4741 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4801 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4861 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4921 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4981 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5041 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5101 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5161 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5221 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5281 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5341 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5401 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5461 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5521 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5581 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6001 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6241 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6961 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7561 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     7621 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7681 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7741 gacaattgca tgaagaatct gcttagg
//
