LOCUS       V032686                 6804 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032686
VERSION     V032686
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..1484
                     /label="Vapb(NM_019806)"
                     /note="Vapb(NM_019806)"
                     /gene="Vapb"
     CDS             1491..1556
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1580..2168
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2234..2282
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2484..2912
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2926..3255
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3322..4116
                     /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    4292..4413
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4462..4478)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4486..4502
                     /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(4510..4540)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4555..4576
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4864..5452)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5623..6483)
                     /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(6484..6588)
                     /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 gaaggtggaa caggtcctga
      781 gcctcgagcc acaacacgag ctcaagttcc gaggtccctt cactgatgtt gtcaccacca
      841 acctaaagct tggcaaccca acagaccgaa atgtgtgttt taaagtgaag accacagcac
      901 ctcgcaggta ctgcgtgcgg cccaacagtg gggtcattga tgccggggcc tctctcaatg
      961 tgtctgtgat gttacagcct ttcgattatg atcccaatga gaaaagtaaa cacaagttta
     1021 tggttcagtc tatgtttgct ccgcctgaca cttctgatat ggaggcagta tggaaggagg
     1081 caaaaccgga agaccttatg gattcaaaac ttagatgtgt gtttgaattg ccagcagaaa
     1141 atgctaaacc acatgatgta gaaataaata aaatcatacc tacgagtgca tccaagacag
     1201 aagcaccggc agccgcaaag tccctgacat cgcccctcga tgacacagaa gtaaagaagg
     1261 tgatggaaga gtgcaggcgg ctgcaggggg aggtgcagag gcttcgggag gagagcaggc
     1321 agctcaagga agaagacgga cttcgggtga ggaaggcgat gccgagcaac agccccgtgg
     1381 cggctctggc ggccactggg aaggaggagg gcctgagcgc ccggctgctg gccctggtgg
     1441 ttctgttctt tatcgttggt gtcattatag ggaagattgc cttgggatcc gactacaaag
     1501 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     1561 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1621 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1681 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     1741 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     1801 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     1861 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     1921 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     1981 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2041 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2101 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2161 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     2221 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     2281 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     2341 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     2401 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     2461 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2521 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2581 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2641 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2701 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2761 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2821 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     2881 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     2941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3001 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3061 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3121 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3181 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3241 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3301 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3361 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3421 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3481 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3541 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3601 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3661 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3721 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3781 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3841 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3901 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3961 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4021 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4081 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     4141 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4201 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4261 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4381 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4441 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4501 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4561 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4621 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5041 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5281 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6601 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     6661 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     6721 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     6781 aattgcatga agaatctgct tagg
//
