LOCUS       V023442                 8075 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023442
VERSION     V023442
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             941..3565
                     /label="ENPP3(NM_005021)"
                     /note="ENPP3(NM_005021)"
                     /gene="ENPP3"
     CDS             3572..3637
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3675..3899
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3945..4373
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4387..4716
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4783..5577
                     /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    5751..5872
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5921..5937)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5945..5961
                     /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(5969..5999)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6014..6035
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6323..6908)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7079..7939)
                     /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(7940..8044)
                     /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 atccgccacc atggaatcta cgttgacttt
      961 agcaacggaa caacctgtta agaagaacac tcttaagaaa tataaaatag cttgcattgt
     1021 tcttcttgct ttgctggtga tcatgtcact tggattaggc ctggggcttg gactcaggaa
     1081 actggaaaag caaggcagct gcaggaagaa gtgctttgat gcatcattta gaggactgga
     1141 gaactgccgg tgtgatgtgg catgtaaaga ccgaggtgat tgctgctggg attttgaaga
     1201 cacctgtgtg gaatcaactc gaatatggat gtgcaataaa tttcgttgtg gagagaccag
     1261 attagaggcc agcctttgct cttgttcaga tgactgtttg cagaggaaag attgctgtgc
     1321 tgactataag agtgtttgcc aaggagaaac ctcatggctg gaagaaaact gtgacacagc
     1381 ccagcagtct cagtgcccag aagggtttga cctgccacca gttatcttgt tttctatgga
     1441 tggatttaga gctgaatatt tatacacatg ggatacttta atgccaaata tcaataaact
     1501 gaaaacatgt ggaattcatt caaaatacat gagagctatg tatcctacca aaaccttccc
     1561 aaatcattac accattgtca cgggcttgta tccagagtca catggcatca ttgacaataa
     1621 tatgtatgat gtaaatctca acaagaattt ttcactttct tcaaaggaac aaaataatcc
     1681 agcctggtgg catgggcaac caatgtggct gacagcaatg tatcaaggtt taaaagccgc
     1741 tacctacttt tggcccggat cagaagtggc tataaatggc tcctttcctt ccatatacat
     1801 gccttacaac ggaagtgtcc catttgaaga gaggatttct acactgttaa aatggctgga
     1861 cctgcccaaa gctgaaagac ccaggtttta taccatgtat tttgaagaac ctgattcctc
     1921 tggacatgca ggtggaccag tcagtgccag agtaattaaa gccttacagg tagtagatca
     1981 tgcttttggg atgttgatgg aaggcctgaa gcagcggaat ttgcacaact gtgtcaatat
     2041 catccttctg gctgaccatg gaatggacca gacttattgt aacaagatgg aatacatgac
     2101 tgattatttt cccagaataa acttcttcta catgtacgaa gggcctgccc cccgcatccg
     2161 agctcataat atacctcatg acttttttag ttttaattct gaggaaattg ttagaaacct
     2221 cagttgccga aaacctgatc agcatttcaa gccctatttg actcctgatt tgccaaagcg
     2281 actgcactat gccaagaacg tcagaatcga caaagttcat ctctttgtgg atcaacagtg
     2341 gctggctgtt aggagtaaat caaatacaaa ttgtggagga ggcaaccatg gttataacaa
     2401 tgagtttagg agcatggagg ctatctttct ggcacatgga cccagtttta aagagaagac
     2461 tgaagttgaa ccatttgaaa atattgaagt ctataaccta atgtgtgatc ttctacgcat
     2521 tcaaccagca ccaaacaatg gaacccatgg tagtttaaac catcttctga aggtgccttt
     2581 ttatgagcca tcccatgcag aggaggtgtc aaagttttct gtttgtggct ttgctaatcc
     2641 attgcccaca gagtctcttg actgtttctg ccctcaccta caaaatagta ctcagctgga
     2701 acaagtgaat cagatgctaa atctcaccca agaagaaata acagcaacag tgaaagtaaa
     2761 tttgccattt gggaggccta gggtactgca gaagaacgtg gaccactgtc tcctttacca
     2821 cagggaatat gtcagtggat ttggaaaagc tatgaggatg cccatgtgga gttcatacac
     2881 agtcccccag ttgggagaca catcgcctct gcctcccact gtcccagact gtctgcgggc
     2941 tgatgtcagg gttcctcctt ctgagagcca aaaatgttcc ttctatttag cagacaagaa
     3001 tatcacccac ggcttcctct atcctcctgc cagcaataga acatcagata gccaatatga
     3061 tgctttaatt actagcaatt tggtacctat gtatgaagaa ttcagaaaaa tgtgggacta
     3121 cttccacagt gttcttctta taaaacatgc cacagaaaga aatggagtaa atgtggttag
     3181 tggaccaata tttgattata attatgatgg ccattttgat gctccagatg aaattaccaa
     3241 acatttagcc aacactgatg ttcccatccc aacacactac tttgtggtgc tgaccagttg
     3301 taaaaacaag agccacacac cggaaaactg ccctgggtgg ctggatgtcc taccctttat
     3361 catccctcac cgacctacca acgtggagag ctgtcctgaa ggtaaaccag aagctctttg
     3421 ggttgaagaa agatttacag ctcacattgc ccgggtccgt gatgtagaac ttctcactgg
     3481 gcttgacttc tatcaggata aagtgcagcc tgtctctgaa attttgcaac taaagacata
     3541 tttaccaaca tttgaaacca ctattctcga ggactacaaa gaccatgacg gtgattataa
     3601 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg
     3661 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3721 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3781 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3841 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3901 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4621 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4681 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4741 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5281 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5581 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5641 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5701 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5761 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5821 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5881 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6061 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6121 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6241 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6301 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6361 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6421 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6481 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6541 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6601 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6661 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6721 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6781 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6841 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6961 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7021 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7081 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7141 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7201 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7261 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7321 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7381 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7441 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7501 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7561 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7621 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7681 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7741 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7801 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7861 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7921 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7981 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8041 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
