LOCUS       V034629                 7387 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034629
VERSION     V034629
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"
     CDS             964..2877
                     /label="GBP7(NM_207398)"
                     /note="GBP7(NM_207398)"
                     /gene="GBP7"
     CDS             2884..2949
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2987..3211
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3257..3685
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3699..4028
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4095..4889
                     /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    5063..5184
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5233..5249)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5257..5273
                     /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(5281..5311)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5326..5347
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5635..6220)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6391..7251)
                     /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(7252..7356)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcat cagagatcca catgccaggc ccagtgtgcc tcactgagaa cactaaaggg
     1021 catctggtgg tgaattcaga agctctggaa atcctgtctg ccattacaca gcctgtagta
     1081 gtggtggcaa ttgtgggcct ctaccgcaca ggcaaatcct acctaatgaa caagctggct
     1141 gggaagaaca aaggcttccc tctgggctgc acagtgaagt ctgaaaccaa aggcatctgg
     1201 atgtggtgtg tgccccaccc ctccaagcca aaccacaccc tgatccttct ggacacggag
     1261 ggcctgggtg atatggaaaa gagtgaccct aagagtgact cgtggatctt tgccctggct
     1321 gtgcttctaa gcagcagctt tgtctacaac agcatgggca ccatcaacca ccaggccctg
     1381 gagcagctgc actacgtgac tgagctaaca gagctaatca gggcaaaatc gtgccccaga
     1441 cctgatgaag ttgaggactc cagcgagttt gtgagtttct ttccagactt tatttggact
     1501 gttcgagatt ttaccctgga gctgaagtta gatggacacc ccatcacaga agatgagtac
     1561 ctggagaatg ccttgaagct gatttcaggc aagaatcccc aaatccaaaa ttctaacaag
     1621 cccagggagt ggatcaggca tttctttcca aaacagaagt gctttgtctt tgaccggcca
     1681 ataaatgaca aaaaactctt actccatgtt gaagaagtac gagaagacca actggatagt
     1741 aatttccaga tgcaatcaga aaatttctgt tcttatatct tcacccatgc aaagaccaag
     1801 accctgagag agggaatcct tgtcactgga aaccggctgg ggatgctggt ggagacctac
     1861 ctggatgcca tcaacagtgg agcgactcct tgtctggaga atgcaatggc agttctggcc
     1921 cagtgtgaga actcagcagc cgtgcagagg gcagccaacc actacagcca gcagatggcc
     1981 cagcaagtga gattccccac agacacactc caggagctgc tggacgtgca tgcagtttgt
     2041 gagagggaag ccattgcagt cttcatggag tactccttca aagataaaag ccaggaattt
     2101 cagaagaagc ttgtggacac catggagaaa aagaaggaag actttgtgct gcagaatgaa
     2161 gaggcatctg ccaaatattg tcaggctgag cttaagcggc tttcagagct cttgacagaa
     2221 agtatttcaa gaggaacttt ctttgttccg ggggggcaca atatctactt agaagcaaaa
     2281 aagaagattg aacaggacta tacactagtg cccagaaaag gagttaaggc agacgaggtc
     2341 ctccagagct tcctgcagtc acaggtggtt atagaggaat ccatcctgca gtcagacaaa
     2401 gccctcactg ctggagagaa ggccatagca gctaagcagg ctaagaagga ggcagctgaa
     2461 aaggaacagg agctgctaag acaaaaacag aaggaacagc agcaaatgat ggaggctcaa
     2521 gagagaagtt tccaggaaaa catagctcaa ctcaagaaga agatggagag ggaaagggaa
     2581 aactatatga gagaactgag aaagatgttg agtcacaaga tgaaggtcct agaagaactg
     2641 cttactgaag gatttaaaga gatatttgag tcgttaaatg aagagattaa tcgactgaaa
     2701 gaacaaattg aagcagctga aaatgaagag ccctcagtgt tttcacagat tcttgatgtg
     2761 gctggcagta tatttattgc agcactacct ggggctgcta agctagttga tttaggaatg
     2821 aaaattctta gctcattatg taataggctg agaaatcctg gtaagaaaat tataagcctc
     2881 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     2941 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3001 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3061 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3121 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3181 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3241 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3301 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3361 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3421 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3481 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3541 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3601 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3661 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3721 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3781 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3841 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3901 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3961 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4021 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4081 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4141 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4201 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4261 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4321 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4381 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4441 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4501 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4561 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4621 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4681 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4741 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4801 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4861 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4921 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4981 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5041 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5101 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5161 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5221 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5281 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5341 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5401 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5461 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5521 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5581 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5641 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5701 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5761 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5821 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5881 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5941 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6001 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6061 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6121 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6181 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6241 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6301 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6361 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6421 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6481 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6541 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6601 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6661 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6721 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6781 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6841 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6901 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6961 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7021 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7081 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7141 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7201 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7381 tgacgtc
//
