LOCUS       V019615                 7304 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019615
VERSION     V019615
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..1984
                     /label="ENO2(NM_001975)"
                     /note="ENO2(NM_001975)"
                     /gene="ENO2"
     CDS             1991..2056
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2080..2668
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2734..2782
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2984..3412
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3426..3755
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3822..4616
                     /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    4792..4913
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4962..4978)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4986..5002
                     /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(5010..5040)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5055..5076
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5364..5952)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6123..6983)
                     /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(6984..7088)
                     /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 ccatgtccat agagaagatc tgggcccggg agatcctgga
      721 ctcccgcggg aaccccacag tggaggtgga tctctatact gccaaaggtc ttttccgggc
      781 tgcagtgccc agtggagcct ctacgggcat ctatgaggcc ctggagctga gggatggaga
      841 caaacagcgt tacttaggca aaggtgtcct gaaggcagtg gaccacatca actccaccat
      901 cgcgccagcc ctcatcagct caggtctctc tgtggtggag caagagaaac tggacaacct
      961 gatgctggag ttggatggga ctgagaacaa atccaagttt ggggccaatg ccatcctggg
     1021 tgtgtctctg gccgtgtgta aggcaggggc agctgagcgg gaactgcccc tgtatcgcca
     1081 cattgctcag ctggccggga actcagacct catcctgcct gtgccggcct tcaacgtgat
     1141 caatggtggc tctcatgctg gcaacaagct ggccatgcag gagttcatga tcctcccagt
     1201 gggagctgag agctttcggg atgccatgcg actaggtgca gaggtctacc atacactcaa
     1261 gggagtcatc aaggacaaat acggcaagga tgccaccaat gtgggggatg aaggtggctt
     1321 tgcccccaat atcctggaga acagtgaagc cttggagctg gtgaaggaag ccatcgacaa
     1381 ggctggctac acggaaaaga tcgttattgg catggatgtt gctgcctcag agttttatcg
     1441 tgatggcaaa tatgacttgg acttcaagtc tcccactgat ccttcccgat acatcactgg
     1501 ggaccagctg ggggcactct accaggactt tgtcagggac tatcctgtgg tctccattga
     1561 ggacccattt gaccaggatg attgggctgc ctggtccaag ttcacagcca atgtagggat
     1621 ccagattgtg ggtgatgacc tgacagtgac caacccaaaa cgtattgagc gggcagtgga
     1681 agaaaaggcc tgcaactgtc tgctgctcaa ggtcaaccag atcggctcgg tcactgaagc
     1741 catccaagcg tgcaagctgg cccaggagaa tggctggggg gtcatggtga gtcatcgctc
     1801 aggagagact gaggacacat tcattgctga cctggtggtg gggctgtgca caggccagat
     1861 caagactggt gccccgtgcc gttctgaacg tctggctaaa tacaaccagc tcatgagaat
     1921 tgaggaagag ctgggggatg aagctcgctt tgccggacat aacttccgta atcccagtgt
     1981 gctgctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     2041 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     2101 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2161 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2221 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2281 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2341 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2401 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2461 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     2521 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2581 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2641 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     2701 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2761 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     2821 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     2881 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     2941 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3001 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3061 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3121 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3181 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3241 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3301 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     3361 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3661 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3721 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3781 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3841 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3901 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3961 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4021 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4081 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4141 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4201 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4261 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4321 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4381 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4441 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4501 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4561 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4621 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4681 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4741 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4801 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4861 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4921 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4981 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5041 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5101 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5161 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5221 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5281 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5341 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5401 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5461 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5521 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5581 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5641 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5701 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5761 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5821 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5881 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5941 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6001 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6061 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6121 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6181 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6241 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6301 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6361 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6421 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6481 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6541 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6601 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6661 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6721 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6781 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6841 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6901 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6961 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7021 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7081 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7141 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     7201 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     7261 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
