LOCUS       V043640                 6688 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043640
VERSION     V043640
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2208
                     /label="VMP1(NM_030938)"
                     /note="VMP1(NM_030938)"
                     /gene="VMP1"
     polyA_signal    2288..2512
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2558..2986
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3000..3329
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3396..4190
                     /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    4364..4485
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4534..4550)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4558..4574
                     /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(4582..4612)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4627..4648
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4936..5521)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5692..6552)
                     /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(6553..6657)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggcagaga atggaaaaaa ttgtgaccag
     1021 agacgtgtag caatgaacaa ggaacatcat aatggaaatt tcacagaccc ctcttcagtg
     1081 aatgaaaaga agaggaggga gcgggaagaa aggcagaata ttgtcctgtg gagacagccg
     1141 ctcattacct tgcagtattt ttctctggaa atccttgtaa tcttgaagga atggacctca
     1201 aaattatggc atcgtcaaag cattgtggtg tcttttttac tgctgcttgc tgtgcttata
     1261 gctacgtatt atgttgaagg agtgcatcaa cagtatgtgc aacgtataga gaaacagttt
     1321 cttttgtatg cctactggat aggcttagga attttgtctt ctgttgggct tggaacaggg
     1381 ctgcacacct ttctgcttta tctgggtcca catatagcct cagttacatt agctgcttat
     1441 gaatgcaatt cagttaattt tcccgaacca ccctatcctg atcagattat ttgtccagat
     1501 gaagagggca ctgaaggaac catttctttg tggagtatca tctcaaaagt taggattgaa
     1561 gcctgcatgt ggggtatcgg tacagcaatc ggagagctgc ctccatattt catggccaga
     1621 gcagctcgcc tctcaggtgc tgaaccagat gatgaagagt atcaggaatt tgaagagatg
     1681 ctggaacatg cagagtctgc acaagacttt gcctcccggg ccaaactggc agttcaaaaa
     1741 ctagtacaga aagttggatt ttttggaatt ttggcctgtg cttcaattcc aaatccttta
     1801 tttgatctgg ctggaataac gtgtggacac tttctggtac ctttttggac cttctttggt
     1861 gcaaccctaa ttggaaaagc aataataaaa atgcatatcc agaaaatttt tgttataata
     1921 acattcagca agcacatagt ggagcaaatg gtggctttca ttggtgctgt ccccggcata
     1981 ggtccatctc tgcagaagcc atttcaggag tacctggagg ctcaacggca gaagcttcac
     2041 cacaaaagcg aaatgggcac accacaggga gaaaactggt tgtcctggat gtttgaaaag
     2101 ttggtcgttg tcatggtgtg ttacttcatc ctatctatca ttaactccat ggcacaaagt
     2161 tatgccaaac gaatccagca gcggttgaac tcagaggaga aaactaaata agaattctgc
     2221 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     2281 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2341 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2401 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2461 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2521 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2581 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2641 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2701 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2761 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2821 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2881 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2941 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3001 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3061 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3121 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3181 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3241 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3301 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3361 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3421 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3481 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3541 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3601 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3661 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3721 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3781 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3841 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3901 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3961 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4021 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4081 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4141 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4201 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4261 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4321 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4381 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4441 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4501 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4561 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4621 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4681 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4741 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4801 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4861 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4921 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4981 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5041 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5101 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5161 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5221 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5281 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5341 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5401 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5461 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5521 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5581 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5641 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5701 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5761 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5821 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5881 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5941 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6001 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6061 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6121 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6181 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6241 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6301 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6361 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6421 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6481 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6541 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6601 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6661 atttccccga aaagtgccac ctgacgtc
//
