LOCUS       V038893                 8574 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038893
VERSION     V038893
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..3254
                     /label="Veph1(NM_145820)"
                     /note="Veph1(NM_145820)"
                     /gene="Veph1"
     CDS             3261..3326
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3350..3938
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4004..4052
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4254..4682
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4696..5025
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5092..5886
                     /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    6062..6183
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6232..6248)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6256..6272
                     /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(6280..6310)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6325..6346
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6634..7222)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7393..8253)
                     /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(8254..8358)
                     /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 ccaccatgca tcaacttttc agattggttt
      781 tgggacaaaa agatctttca aaagctgggg acctcttctc tttagatgat gctgagattg
      841 aagacagcct gacagaagct ttggagcaga ttaaagtaat tagctcatct ttggattacc
      901 aaactaataa caatgaccag gcggtggttg aaatctgtat cacaaggatt accacagcca
      961 tcagagagac agaatccatt gaaaagcatg ccagagccct ggtggggctg tgggattcct
     1021 gcttggaaca caacctgaga cctgcaggga aagacgaaga tactccacat gcaaaaatcg
     1081 cctctgatat catgagttgc atcttacaga attacaaccg gacccctgtg atggtcttag
     1141 ctgtccccat cgcagtgaaa ttcctccaca gaggcagcaa ggagctgtgt aggaacatgt
     1201 ccaactacct gtccctggct gccatcacca aggcggatct gctggctgac cacactgaag
     1261 ggataataaa gagcatactc caaggaaatg ccatgctgtt aagggtgcta cctgctgtat
     1321 atgaaaagca gccccaacca atcaacagac acctggcaga actcctggcc ttgatgtctc
     1381 aattggagca gacagaacag taccatctac taaggcttct gcacgtagca gcaaagagaa
     1441 aagatgtgga ggtggttcag aaatgtgttc ctttcctgat cagaaatttg aaggattcaa
     1501 cctataatga catcatccta aacattctca tagaaatagc aggccatgag ccactggcct
     1561 tgaacagttt cctcccgatg ctgaaagaga ttgcagagca attcccctac ctcactggac
     1621 agatggcaag gatctttggt gctgttggtc atgtggatga agagagagcc aggagctgcc
     1681 tacgatacct ggtgagccag ctggctaata tggagcatcc attccaccat attctcctgc
     1741 tggaaattaa gagtatcaca gatgccttct cctccatcct gggcccgcac agcagggaca
     1801 tcttccgaat gagcaacagc ttcaccaaca ttgccaagct cctttccaga caactggaaa
     1861 acagcaaggc tgacagtagc aggagaaaaa ccagcactga agttagcatt cctgagaaac
     1921 tgagagaact caattccatg gagcctgaaa gtgaagacca tgaaaaactc caagttaaaa
     1981 tccaagcctt tgaagacaaa ataaatgctg agagcaatac cccaggctct ggccggagat
     2041 acagcttaga ccacatctct aaggaagaac gaaaaagcat tagatttagc aggtccagaa
     2101 gtttggcttt gaatactgtg ctcacaaacg gtgtgagtgt ggaagataac gaagtcgaag
     2161 aaaaggcagg aatgcacgcc agcatctctc tttcgcaaat agacccactc agccacggga
     2221 ttggcaagtt gccctttaag acagacactc atggatcacc actgagaaat tcctcagctt
     2281 cacacccaag tattatacat actgaaccag agactatgcc cgaaactttt aaagaaaaca
     2341 tccaagaaga aattctagag gcagccacaa gtcctattga ataccaagat aagctctact
     2401 tgcatttaag ggaaaacctc agcaaagtga aggcttatgc cttggaaatc gcaaaaaagg
     2461 tcccaatccc agaccagtgc accattgaag atactatgag aagttgcgtg gcaaagctgt
     2521 tctttacctg ttctttaaag ggccattact gcctatacag caagtccagc ttcatccttg
     2581 tcagccaagc accccagccg tggatccagg tcatgtttct ctcccaacag agcctgtttc
     2641 ctgagcccct gtccattcag agtgggtctg tgcagttcct caaagcccta tgggagaaaa
     2701 cgcaggacac gggagcgcac agctttgaag tcgccatgac agagtccacc tttccacagc
     2761 agaaggatct ggagcagctt cagctccatc tggaagaagt gcggttcttc gacgtgtttg
     2821 gcttcagtga aacagcgggg gcatggcagt gcttcatgtg caacaaccct gaaaaggcaa
     2881 ctgttgtaaa tcaagatggc cagcctctca tcgaaggaaa acttaaggag aaacaagtga
     2941 ggtggaagtt catcaagagg tggaaaacgc attattttac actggccgga aaccaacttc
     3001 tgtttcaaaa ggggaaatct aaggatgacc ctgatgattc cccgatagaa ctcagcaagg
     3061 tacagagcgt gaaagctgtg gctaagaaac gccgggaccg ctctctgcct cgggcgtttg
     3121 aaatcttcac agatagtaag acctatgttt tcaaagccaa ggatgagaag aacgcagagg
     3181 aatggctgca gtgcatcaac gtggcgctgg cccaggcaaa ggagagggaa agccgagaag
     3241 taaccacata tctgggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     3301 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3361 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3421 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3481 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3541 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3601 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3661 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3721 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3781 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3841 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3901 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3961 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4021 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4081 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4141 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4201 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     4261 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4321 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4381 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4441 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4501 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4561 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4621 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4681 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4741 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4801 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4861 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4921 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4981 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5041 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     5101 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5161 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5221 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5281 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5341 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5401 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5461 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5521 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5581 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5641 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5701 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5761 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5821 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5881 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5941 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6001 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6061 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6121 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6181 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6241 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6301 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6361 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6421 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7681 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7981 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8401 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8461 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8521 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
