LOCUS       V038527                 7110 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038527
VERSION     V038527
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..1790
                     /label="Gmpr(NM_025508)"
                     /note="Gmpr(NM_025508)"
                     /gene="Gmpr"
     CDS             1797..1862
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1886..2474
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2540..2588
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2790..3218
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3232..3561
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3628..4422
                     /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    4598..4719
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4768..4784)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4792..4808
                     /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(4816..4846)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4861..4882
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5170..5758)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5929..6789)
                     /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(6790..6894)
                     /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 ccaccatgcc ccgcatagac gcggacctta
      781 aactcgactt caaggatgtt ctgctgcgac ctaagcggag cagcctcaag agccgatccg
      841 aggtggatct tgagcgaact tttacatttc gaaactcgaa gcaaacctac tcagggattc
      901 ccatcattgt ggccaacatg gacactgtgg ggacatttga gatggctgtg gtgatgtcac
      961 agcatgccat gtttacagcc gttcacaagc actactccct ggatgactgg aaatgttttg
     1021 cggaaaccca ccccgaatgc ctgcagcatg tagccgtgag ttctggcagc gggcagaatg
     1081 atcttgagag gatgagccgc atcttggaag ctgtgcctca ggtgaagttc atctgcctgg
     1141 atgtggccaa tgggtattca gagcattttg tggaattcgt gaaactggtc cgatccaaat
     1201 tccccgaaca caccatcatg gcagggaacg tggtgaccgg agagatggtg gaagagctca
     1261 tcctctctgg agcagatatc atcaaagtgg gagtcggacc aggttctgtg tgcaccaccc
     1321 gaaccaagac aggagtgggc tacccgcagc tgagtgcggt gatagagtgt gctgactccg
     1381 cccatggcct caaggggcac atcatctctg atggaggctg cacatgtcca ggagatgtcg
     1441 ccaaagcctt tggagctggt gcagactttg tcatgctggg aggcatgttt tcaggccaca
     1501 ccgagtgtgc aggtgaggtg atcgagagga acgggcagaa gctgaaactc ttctatggca
     1561 tgagctcaga cacagccatg aagaaacacg caggaggcgt cgccgagtac agggcctccg
     1621 agggcaagac cgtggaagtg ccttacaaag gggacgtgga gaacaccatt ctggatatcc
     1681 ttggaggact gcgctccacc tgtacctacg tcggggctgc caagctcaaa gagctcagca
     1741 ggagagccac gttcatccgg gtgacccaac agcacaacac ggtgtttggt ggatccgact
     1801 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     1861 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     1921 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1981 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2041 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2101 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2161 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2221 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2281 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2341 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2401 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2461 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     2521 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     2581 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     2641 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     2701 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     2761 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2821 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2881 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2941 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3001 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3061 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3121 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     3181 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3241 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3361 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3481 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3541 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3601 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3661 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3721 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3781 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3841 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3901 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3961 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4021 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4081 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4141 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4201 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4261 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4321 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4381 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4441 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4501 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4561 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4621 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4681 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4741 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4801 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4861 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4921 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4981 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5041 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5101 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5161 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5221 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5281 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5341 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5401 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5461 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5521 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5581 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5641 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5701 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5761 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5821 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5881 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6001 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6061 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6541 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6781 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6841 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6901 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     6961 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7021 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7081 accgacaatt gcatgaagaa tctgcttagg
//
