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 //