LOCUS       V035651                 8218 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035651
VERSION     V035651
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"
     CDS             747..2120
                     /label="ENPP7(NM_178543)"
                     /note="ENPP7(NM_178543)"
                     /gene="ENPP7"
     CDS             2127..2216
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             2253..2963
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    2994..3582
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3648..3696
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3898..4326
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4340..4669
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4736..5530
                     /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    5706..5827
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5876..5892)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5900..5916
                     /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(5924..5954)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5969..5990
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6278..6866)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7037..7897)
                     /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(7898..8002)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagaaa gcttggtacc gccaccatga gaggcccggc cgtcctcctc actgtggctc
      781 tggccacgct cctggctccc ggggccggag caccggtaca aagtcagggc tcccagaaca
      841 agctgctcct ggtgtccttc gacggcttcc gctggaacta cgaccaggac gtggacaccc
      901 ccaacctgga cgccatggcc cgagacgggg tgaaggcacg ctacatgacc cccgcctttg
      961 tcaccatgac cagcccctgc cacttcaccc tggtcaccgg caaatatatc gagaaccacg
     1021 gggtggttca caacatgtac tacaacacca ccagcaaggt gaagctgccc taccacgcca
     1081 cgctgggcat ccagaggtgg tgggacaacg gcagcgtgcc catctggatc acagcccaga
     1141 ggcagggcct gagggctggc tccttcttct acccgggcgg gaacgtcacc taccaagggg
     1201 tggctgtgac gcggagccgg aaagaaggca tcgcacacaa ctacaaaaat gagacggagt
     1261 ggagagcgaa catcgacaca gtgatggcgt ggttcacaga ggaggacctg gatctggtca
     1321 cactctactt cggggagccg gactccacgg gccacaggta cggccccgag tccccggaga
     1381 ggagggagat ggtgcggcag gtggaccgga ccgtgggcta cctccgggag agcatcgcgc
     1441 gcaaccacct cacagaccgc ctcaacctga tcatcacatc cgaccacggc atgacgaccg
     1501 tggacaaacg ggctggcgac ctggttgaat tccacaagtt ccccaacttc accttccggg
     1561 acatcgagtt tgagctcctg gactacggac caaacgggat gctgctccct aaagaaggga
     1621 ggctggagaa ggtgtacgat gccctcaagg acgcccaccc caagctccac gtctacaaga
     1681 aggaggcgtt ccccgaggcc ttccactacg ccaacaaccc cagggtcaca cccctgctga
     1741 tgtacagcga ccttggctac gtcatccatg ggagaattaa cgtccagttc aacaatgggg
     1801 agcacggctt tgacaacaag gacatggaca tgaagaccat cttccgcgct gtgggcccta
     1861 gcttcagggc gggcctggag gtggagccct ttgagagcgt ccacgtgtac gagctcatgt
     1921 gccggctgct gggcatcgtg cccgaggcca acgatgggca cctagctact ctgctgccca
     1981 tgctgcacac agaatctgct cttccgcctg atggaaggcc tactctcctg cccaagggaa
     2041 gatctgctct cccgcccagc agcaggcccc tcctcgtgat gggactgctg gggaccgtga
     2101 ttcttctgtc tgaggtcgca ctcgaggaac aaaaactaat atcagaggaa gatttggaac
     2161 agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag gacttaggtg
     2221 gaggcggatc aggaggcggt ggatctggat ccatggtgag caagggcgag gaggataaca
     2281 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     2341 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2401 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     2461 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     2521 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2581 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     2641 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     2701 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     2761 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     2821 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     2881 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     2941 gcatggacga gctgtacaag tagaccggtt agtaatgagt ttgatatctc gacaatcaac
     3001 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3061 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3121 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3181 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3241 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3301 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3361 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3421 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3481 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3541 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct
     3601 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     3661 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     3721 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     3781 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     3841 gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg
     3901 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3961 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4021 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     4081 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4141 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4201 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4261 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4321 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4381 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4441 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4501 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4561 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4621 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4681 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4741 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4801 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4861 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4921 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4981 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5041 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5101 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5161 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5221 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5281 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5341 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5401 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5461 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5521 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     5581 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5641 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5701 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5821 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5881 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5941 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6001 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6061 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6121 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6181 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6241 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6301 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6361 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6421 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     6481 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6541 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6601 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6661 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6721 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6781 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6841 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6901 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6961 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7021 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7081 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7141 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7201 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7261 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7321 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7381 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7441 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7501 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7561 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7621 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7681 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7741 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7801 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7861 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7921 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7981 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8041 cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata
     8101 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8161 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
