LOCUS       V033623                 7003 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033623
VERSION     V033623
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..2493
                     /label="NFIX(NM_001271043)"
                     /note="NFIX(NM_001271043)"
                     /gene="NFIX"
     CDS             2500..2565
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2603..2827
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2873..3301
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3315..3644
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3711..4505
                     /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    4679..4800
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4849..4865)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4873..4889
                     /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(4897..4927)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4942..4963
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5251..5836)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6007..6867)
                     /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(6868..6972)
                     /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 accatggaga tgtcatgggc gcgacagagc ctggcgggga taccagcagc gtgtgatgag
     1021 ttccacccgt tcatcgaggc actgctgcct cacgtccgcg ctttctccta cacctggttc
     1081 aacctgcagg cgcggaagcg caagtacttc aagaagcatg aaaagcggat gtcgaaggac
     1141 gaggagcggg cggtgaagga cgagctgctg ggcgagaagc ccgagatcaa gcagaagtgg
     1201 gcatcccggc tgctggccaa gctgcgcaag gacatccggc ccgagttccg cgaggacttc
     1261 gtgctgacca tcacgggcaa gaagcccccc tgctgcgtgc tctccaaccc cgaccagaag
     1321 ggcaagatcc ggcggattga ctgcctgcgc caggctgaca aggtgtggcg gctggacctg
     1381 gtcatggtga ttttgtttaa ggggatcccc ctggaaagta ctgatgggga gcggctctac
     1441 aagtcgcctc agtgctcgaa ccccggcctg tgcgtccagc cacatcacat tggagtcaca
     1501 atcaaagaac tggatcttta tctggcttac tttgtccaca ctccggaatc cggacaatca
     1561 gatagttcaa accagcaagg agatgcggac atcaaaccac tgcccaacgg gcacttaagt
     1621 ttccaggact gttttgtgac ttccggggtc tggaatgtga cggagctggt gagagtatca
     1681 cagactcctg ttgcaacagc atcagggccc aacttctccc tggcggacct ggagagtccc
     1741 agctactaca acatcaacca ggtgaccctg gggcggcggt ccatcacctc ccctccttcc
     1801 accagcacca ccaagcgccc caagtccatc gatgacagtg agatggagag ccctgttgat
     1861 gacgtgttct atcccgggac aggccgttcc ccagcagctg gcagcagcca gtccagcggg
     1921 tggcccaacg atgtggatgc aggcccggct tctctaaaga agtcaggaaa gctggacttc
     1981 tgcagtgccc tctcctctca gggcagctcc ccgcgcatgg ctttcaccca ccacccgctg
     2041 cctgtgcttg ctggagtcag accagggagc ccccgggcca cagcatcagc cctgcacttc
     2101 ccctccacgt ccatcatcca gcagtcgagc ccgtatttca cgcacccgac catccgctac
     2161 caccaccacc acgggcagga ctcactgaag gagtttgtgc agtttgtgtg ctcggatggc
     2221 tcgggccagg ccaccggaca gcccaacggt agcggccagg gcaaagtccc ggggtcattt
     2281 ttgctaccgc cgccgcctcc agtggccaga cctgtgcccc ttcctatgcc tgattccaaa
     2341 tccaccagca ctgccccaga cggcgccgcc ttgactcctc catcaccttc attcgcaacg
     2401 acaggcgcct cctctgccaa ccggtttgtc agcatcggac cccgggacgg caactttctg
     2461 aacatcccac agcagtctca gtcctggttc ctcctcgagg actacaaaga ccatgacggt
     2521 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgtt
     2581 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2641 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2701 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2761 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2821 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2881 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2941 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3001 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3061 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3121 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3181 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3241 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3301 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3361 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3421 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3481 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3541 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3601 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3661 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3721 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3781 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3841 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3901 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3961 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4021 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4081 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4141 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4201 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4261 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4321 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4381 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4441 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4501 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4561 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4621 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4681 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4741 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4801 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4861 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4921 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4981 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5041 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5101 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5161 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5221 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5281 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5341 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5701 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5761 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     5821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6061 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6121 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6181 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6241 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6301 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6361 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6421 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6481 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6541 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6601 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6661 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6721 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6781 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6841 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6901 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6961 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
