LOCUS V045973 10587 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045973 VERSION V045973 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers promoter 3..229 /note="Rous sarcoma virus enhancer/promoter" /label="RSV promoter" LTR 230..410 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 457..582 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1075..1308 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 1803..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" protein_bind 1938..1958 /bound_moiety="BP Clonase™" /gene="mutant version of attB" /note="core recombination site for the Gateway® BP reaction" /label="attB4" enhancer 2019..2322 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2323..2526 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2566..5190 /label="ENPP3(NM_005021)" /note="ENPP3(NM_005021)" /gene="ENPP3" CDS 5197..5262 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 5302..5890 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" promoter 5921..6420 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6437..7036 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" LTR 7108..7341 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7413..7534 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7574..7709 /note="SV40 origin of replication" /label="SV40 ori" promoter complement(7730..7748) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(7758..7774) /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" rep_origin 7916..8371 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 8397..8501 /gene="bla" /label="AmpR promoter" CDS 8502..9362 /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" rep_origin 9533..10121 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 10409..10430 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 10445..10475 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 10483..10499 /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" primer_bind 10507..10523 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" promoter 10544..10562 /note="promoter for bacteriophage T3 RNA polymerase" /label="T3 promoter" ORIGIN 1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca 61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga 121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt 181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg 241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg 421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt 481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg 541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga 601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta 661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta 721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga 781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg 841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt 901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga 961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc 1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc 1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct 1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag 1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca 1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg 1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa 1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta 1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata 1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt 1921 actagtgatt atcggatcaa ctttgtatag aaaagttgta gttattaata gtaatcaatt 1981 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 2041 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 2101 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 2161 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc 2221 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct 2281 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag 2341 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt 2401 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac 2461 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc 2521 agagctggtt tagtgaaccg tcagatccaa gttggatccg ccaccatgga atctacgttg 2581 actttagcaa cggaacaacc tgttaagaag aacactctta agaaatataa aatagcttgc 2641 attgttcttc ttgctttgct ggtgatcatg tcacttggat taggcctggg gcttggactc 2701 aggaaactgg aaaagcaagg cagctgcagg aagaagtgct ttgatgcatc atttagagga 2761 ctggagaact gccggtgtga tgtggcatgt aaagaccgag gtgattgctg ctgggatttt 2821 gaagacacct gtgtggaatc aactcgaata tggatgtgca ataaatttcg ttgtggagag 2881 accagattag aggccagcct ttgctcttgt tcagatgact gtttgcagag gaaagattgc 2941 tgtgctgact ataagagtgt ttgccaagga gaaacctcat ggctggaaga aaactgtgac 3001 acagcccagc agtctcagtg cccagaaggg tttgacctgc caccagttat cttgttttct 3061 atggatggat ttagagctga atatttatac acatgggata ctttaatgcc aaatatcaat 3121 aaactgaaaa catgtggaat tcattcaaaa tacatgagag ctatgtatcc taccaaaacc 3181 ttcccaaatc attacaccat tgtcacgggc ttgtatccag agtcacatgg catcattgac 3241 aataatatgt atgatgtaaa tctcaacaag aatttttcac tttcttcaaa ggaacaaaat 3301 aatccagcct ggtggcatgg gcaaccaatg tggctgacag caatgtatca aggtttaaaa 3361 gccgctacct acttttggcc cggatcagaa gtggctataa atggctcctt tccttccata 3421 tacatgcctt acaacggaag tgtcccattt gaagagagga tttctacact gttaaaatgg 3481 ctggacctgc ccaaagctga aagacccagg ttttatacca tgtattttga agaacctgat 3541 tcctctggac atgcaggtgg accagtcagt gccagagtaa ttaaagcctt acaggtagta 3601 gatcatgctt ttgggatgtt gatggaaggc ctgaagcagc ggaatttgca caactgtgtc 3661 aatatcatcc ttctggctga ccatggaatg gaccagactt attgtaacaa gatggaatac 3721 atgactgatt attttcccag aataaacttc ttctacatgt acgaagggcc tgccccccgc 3781 atccgagctc ataatatacc tcatgacttt tttagtttta attctgagga aattgttaga 3841 aacctcagtt gccgaaaacc tgatcagcat ttcaagccct atttgactcc tgatttgcca 3901 aagcgactgc actatgccaa gaacgtcaga atcgacaaag ttcatctctt tgtggatcaa 3961 cagtggctgg ctgttaggag taaatcaaat acaaattgtg gaggaggcaa ccatggttat 4021 aacaatgagt ttaggagcat ggaggctatc tttctggcac atggacccag ttttaaagag 4081 aagactgaag ttgaaccatt tgaaaatatt gaagtctata acctaatgtg tgatcttcta 4141 cgcattcaac cagcaccaaa caatggaacc catggtagtt taaaccatct tctgaaggtg 4201 cctttttatg agccatccca tgcagaggag gtgtcaaagt tttctgtttg tggctttgct 4261 aatccattgc ccacagagtc tcttgactgt ttctgccctc acctacaaaa tagtactcag 4321 ctggaacaag tgaatcagat gctaaatctc acccaagaag aaataacagc aacagtgaaa 4381 gtaaatttgc catttgggag gcctagggta ctgcagaaga acgtggacca ctgtctcctt 4441 taccacaggg aatatgtcag tggatttgga aaagctatga ggatgcccat gtggagttca 4501 tacacagtcc cccagttggg agacacatcg cctctgcctc ccactgtccc agactgtctg 4561 cgggctgatg tcagggttcc tccttctgag agccaaaaat gttccttcta tttagcagac 4621 aagaatatca cccacggctt cctctatcct cctgccagca atagaacatc agatagccaa 4681 tatgatgctt taattactag caatttggta cctatgtatg aagaattcag aaaaatgtgg 4741 gactacttcc acagtgttct tcttataaaa catgccacag aaagaaatgg agtaaatgtg 4801 gttagtggac caatatttga ttataattat gatggccatt ttgatgctcc agatgaaatt 4861 accaaacatt tagccaacac tgatgttccc atcccaacac actactttgt ggtgctgacc 4921 agttgtaaaa acaagagcca cacaccggaa aactgccctg ggtggctgga tgtcctaccc 4981 tttatcatcc ctcaccgacc taccaacgtg gagagctgtc ctgaaggtaa accagaagct 5041 ctttgggttg aagaaagatt tacagctcac attgcccggg tccgtgatgt agaacttctc 5101 actgggcttg acttctatca ggataaagtg cagcctgtct ctgaaatttt gcaactaaag 5161 acatatttac caacatttga aaccactatt ctcgaggact acaaagacca tgacggtgat 5221 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtctag atgtacaaag 5281 tggtgataat cgaattccga taatcaacct ctggattaca aaatttgtga aagattgact 5341 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 5401 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 5461 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 5521 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 5581 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 5641 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 5701 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5761 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5821 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 5881 gcctccccgc atcgggaatt cccgcggttc gaattctacc gggtagggga ggcgcttttc 5941 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt 6001 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt 6061 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg 6121 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag 6181 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag 6241 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc 6301 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc 6361 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg 6421 acctcacgtg gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 6481 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 6541 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 6601 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 6661 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 6721 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 6781 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 6841 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 6901 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 6961 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 7021 caagcccggt gcctgaggta cctttaagac caatgactta caaggcagct gtagatctta 7081 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag 7141 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 7201 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 7261 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 7321 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 7381 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 7441 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 7501 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 7561 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 7621 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 7681 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag 7741 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc 7801 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa 7861 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg 7921 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 7981 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 8041 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 8101 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 8161 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 8221 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 8281 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 8341 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg 8401 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 8461 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 8521 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 8581 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 8641 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 8701 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 8761 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 8821 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 8881 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 8941 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 9001 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 9061 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 9121 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 9181 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 9241 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 9301 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 9361 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 9421 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 9481 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 9541 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 9601 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 9661 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 9721 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 9781 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 9841 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 9901 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 9961 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 10021 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 10081 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 10141 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 10201 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc 10261 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac 10321 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact 10381 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc 10441 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat 10501 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa 10561 gggaacaaaa gctggagctg caagctt //