LOCUS       V019707                 7424 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019707
VERSION     V019707
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             683..2104
                     /label="SOX4(NM_003107)"
                     /note="SOX4(NM_003107)"
                     /gene="SOX4"
     CDS             2111..2176
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2200..2788
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2854..2902
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3104..3532
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3546..3875
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3942..4736
                     /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    4912..5033
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5082..5098)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5106..5122
                     /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(5130..5160)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5175..5196
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5484..6072)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6243..7103)
                     /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(7104..7208)
                     /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 ttaagcttgg
      661 taccgagctc ggatccgcca ccatggtgca gcaaaccaac aatgccgaga acacggaagc
      721 gctgctggcc ggcgagagct cggactcggg cgccggcctc gagctgggaa tcgcctcctc
      781 ccccacgccc ggctccaccg cctccacggg cggcaaggcc gacgacccga gctggtgcaa
      841 gaccccgagt gggcacatca agcgacccat gaacgccttc atggtgtggt cgcagatcga
      901 gcggcgcaag atcatggagc agtcgcccga catgcacaac gccgagatct ccaagcggct
      961 gggcaaacgc tggaagctgc tcaaagacag cgacaagatc cctttcattc gagaggcgga
     1021 gcggctgcgc ctcaagcaca tggctgacta ccccgactac aagtaccggc ccaggaagaa
     1081 ggtgaagtcc ggcaacgcca actccagctc ctcggccgcc gcctcctcca agccggggga
     1141 gaagggagac aaggtcggtg gcagtggcgg gggcggccat gggggcggcg gcggcggcgg
     1201 gagcagcaac gcggggggag gaggcggcgg tgcgagtggc ggcggcgcca actccaaacc
     1261 ggcgcagaaa aagagctgcg gctccaaagt ggcgggcggc gcgggcggtg gggttagcaa
     1321 accgcacgcc aagctcatcc tggcaggcgg cggcggcggc gggaaagcag cggctgccgc
     1381 cgccgcctcc ttcgccgccg aacaggcggg ggccgccgcc ctgctgcccc tgggcgccgc
     1441 cgccgaccac cactcgctgt acaaggcgcg gactcccagc gcctcggcct ccgcctcctc
     1501 ggcagcctcg gcctccgcag cgctcgcggc cccgggcaag cacctggcgg agaagaaggt
     1561 gaagcgcgtc tacctgttcg gcggcctggg cacgtcgtcg tcgcccgtgg gcggcgtggg
     1621 cgcgggagcc gaccccagcg accccctggg cctgtacgag gaggagggcg cgggctgctc
     1681 gcccgacgcg cccagcctga gcggccgcag cagcgccgcc tcgtcccccg ccgccggccg
     1741 ctcgcccgcc gaccaccgcg gctacgccag cctgcgcgcc gcctcgcccg ccccgtccag
     1801 cgcgccctcg cacgcgtcct cctcggcctc gtcccactcc tcctcttcct cctcctcggg
     1861 ctcctcgtcc tccgacgacg agttcgaaga cgacctgctc gacctgaacc ccagctcaaa
     1921 ctttgagagc atgtccctgg gcagcttcag ttcgtcgtcg gcgctcgacc gggacctgga
     1981 ttttaacttc gagcccggct ccggctcgca cttcgagttc ccggactact gcacgcccga
     2041 ggtgagcgag atgatctcgg gagactggct cgagtccagc atctccaacc tggttttcac
     2101 ctacctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     2161 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     2221 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2281 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2341 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2401 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2461 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2521 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2581 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     2641 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2701 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2761 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     2821 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2881 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     2941 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3001 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3061 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3121 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3181 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3241 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3301 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3361 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3421 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     3481 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3541 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3601 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3721 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3781 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3841 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3901 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3961 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4021 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4081 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4141 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4201 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4261 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4321 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4381 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4441 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4501 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4561 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4621 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4681 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4741 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4801 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4861 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4921 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4981 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5041 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5101 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5161 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5221 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5281 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5341 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5401 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5461 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5521 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5581 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5641 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5701 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5761 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5821 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5881 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5941 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6001 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6061 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6121 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6181 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6241 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6301 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6361 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6421 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6481 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6541 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6601 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6661 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6721 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6781 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6841 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6901 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6961 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7021 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7081 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7141 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7201 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7261 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     7321 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     7381 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
