LOCUS       V019120                 8234 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019120
VERSION     V019120
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"
     CDS             665..2881
                     /label="SIX5(NM_175875)"
                     /note="SIX5(NM_175875)"
                     /gene="SIX5"
     CDS             2921..2986
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3010..3598
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3664..3712
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3914..4342
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4356..4685
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4752..5546
                     /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    5722..5843
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5892..5908)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5916..5932
                     /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(5940..5970)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5985..6006
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6294..6882)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7053..7913)
                     /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(7914..8018)
                     /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 gctctggcta gcgtttaaac ttaagcttgc
      661 caccatggct accttgcctg cggagccgag cgcggggccg gcggctgggg gggaggcggt
      721 ggcggcggcg gcggcgaccg aagaggagga ggaggaagcg cgccagctct tgcagacttt
      781 gcaggcggcc gagggtgagg cggcggcggc ggccggggcc ggggcgggcg cagcggctgc
      841 gggagctgag ggcccgggat ccccgggcgt ccccgggtcg ccccccgagg ccgcttccga
      901 accgcccacg ggcctccgct tctcgcccga gcaggtggcg tgcgtctgcg aggcgctgct
      961 ccaggcgggc cacgccggcc gcttgagccg cttcctgggc gcactgcccc cggccgagcg
     1021 cctacgtggc agcgacccgg tgttgcgcgc gcgggccctg gtggccttcc agcggggcga
     1081 gtacgccgag ctctaccggc tactcgagag ccgccccttc cccgccgccc accacgcctt
     1141 cctgcaggac ctctacctgc gcgcgcgcta ccatgaggcc gagcgggccc gcggccgcgc
     1201 gcttggcgca gtggacaagt atcgactgcg caagaagttc ccgctgccca agaccatctg
     1261 ggacggcgag gagacagtct actgcttcaa ggagcgctcc cgcgcagcgc tcaaggcctg
     1321 ctaccgcggc aaccgctacc ccacgccgga cgagaagcgc cgcctggcca cactcaccgg
     1381 cctgtcgctc acgcaggtca gcaactggtt caagaaccgg cgacagcgcg accggaccgg
     1441 ggccggaggc ggcgcgccct gcaagagcga gtctgatggg aatcccacga ctgaggacga
     1501 gtccagccga agtcctgagg acctggagag aggggcggcc ccagtgtccg ccgaggccgc
     1561 tgcccagggc tccatattcc tggcagggac cggccctccc gcgccttgcc cggcttcctc
     1621 ctccatcctg gtgaacggga gcttcctggc agccagcggc tccccagcag tgctcctcaa
     1681 cgggggcccc gtcatcatca acggcctggc cctgggcgag gcctccagcc tgggcccgct
     1741 gctgctcact gggggcgggg gtgcccctcc accgcagccc agccctcagg gggccagcga
     1801 gaccaagacc tctctggtcc tggaccctca gacaggggag gtgcggctgg aggaggctca
     1861 gtcggaggcc cctgagacca aaggggccca ggtggctgct ccgggaccag cccttggaga
     1921 ggaggtcctg gggcccctgg cccaagtggt gcctggcccc ccgacggctg ccacctttcc
     1981 tctgcccccg gggccagtgc ctgctgtggc tgccccacaa gtggtaccgc tctccccacc
     2041 cccggggtat cccacgggcc tgagccccac ctccccacta ttgaacctgc cccaggtagt
     2101 acccacctca caggtggtga ccctgcccca ggctgtgggg cccctgcagc tgttggcagc
     2161 cgggccaggc agccctgtga aggtggcagc tgcagcaggc cctgccaatg tgcacctcat
     2221 caactccggg gtgggcgtga ctgccctgca gctgccttcg gccactgccc caggaaactt
     2281 cctcctggcc aaccctgtgt ctggcagccc catcgtgacg ggtgtggccc tgcagcaggg
     2341 caagatcatc ctcaccgcca ccttccccac cagcatgctc gtctcccagg tcctgccgcc
     2401 agcccccggc ctggccctgc cactgaagcc agagacggcc atctccgtgc ctgagggagg
     2461 cctcccggtg gcccccagcc ctgctctccc agaggctcac gccctaggca ccctttctgc
     2521 acagcagcca ccccccgccg ctgccaccac ctccagcacc agcctgccct tctcccctga
     2581 ctcccctggc ctcctgccca acttcccggc gcccccacca gaggggctga tgttgtcacc
     2641 cgcggccgtg cctgtctggt cagcagggct ggaactaagc gcaggaacag aggggctgct
     2701 ggaagcggaa aaggggctgg ggacacaggc cccccacacc gtgctgaggc tgccagaccc
     2761 cgaccctgag gggctgctcc tgggggccac cgcagggggt gaggttgacg aggggttgga
     2821 agctgaggcc aaggttctga cccagctcca gtcggtgcct gtggaggagc ccttggaact
     2881 ggaattctgc agatatccag cacagtggcg gccgctcgag gactacaaag accatgacgg
     2941 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     3001 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3061 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3121 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3181 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3241 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3301 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3361 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     3421 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3481 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3541 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3601 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3661 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3721 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3781 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3841 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3901 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3961 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4021 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     4081 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4141 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4201 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4261 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     4321 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4501 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4561 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4621 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4681 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4741 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4801 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4861 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4921 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4981 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5041 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5101 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5161 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5221 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5281 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5341 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5401 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5461 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5521 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     5581 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5641 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5701 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5761 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5821 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5881 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5941 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6001 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6061 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6121 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6481 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6721 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6901 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6961 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7021 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7081 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7141 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7201 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7261 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7321 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7381 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7441 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7501 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7561 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7621 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7681 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7741 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7801 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7861 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7921 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7981 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8041 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     8101 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     8161 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     8221 agaatctgct tagg
//
