LOCUS V043013 9547 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043013 VERSION V043013 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2843..4243 /label="SMAD2(NM_005901)" /note="SMAD2(NM_005901)" /gene="SMAD2" CDS 4250..4315 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4335..4834 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4855..5454 /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" misc_feature 5468..6056 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6263..6896 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7025..7041) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7049..7065 /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(7073..7103) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7118..7139 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7427..8015) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8186..9046) /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(9047..9151) /gene="bla" /label="AmpR promoter" polyA_signal 9199..9333 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatgtcgtc catcttgcca ttcacgccgc cagttgtgaa 2881 gagactgctg ggatggaaga agtcagctgg tgggtctgga ggagcaggcg gaggagagca 2941 gaatgggcag gaagaaaagt ggtgtgagaa agcagtgaaa agtctggtga agaagctaaa 3001 gaaaacagga cgattagatg agcttgagaa agccatcacc actcaaaact gtaatactaa 3061 atgtgttacc ataccaagca cttgctctga aatttgggga ctgagtacac caaatacgat 3121 agatcagtgg gatacaacag gcctttacag cttctctgaa caaaccaggt ctcttgatgg 3181 tcgtctccag gtatcccatc gaaaaggatt gccacatgtt atatattgcc gattatggcg 3241 ctggcctgat cttcacagtc atcatgaact caaggcaatt gaaaactgcg aatatgcttt 3301 taatcttaaa aaggatgaag tatgtgtaaa cccttaccac tatcagagag ttgagacacc 3361 agttttgcct ccagtattag tgccccgaca caccgagatc ctaacagaac ttccgcctct 3421 ggatgactat actcactcca ttccagaaaa cactaacttc ccagcaggaa ttgagccaca 3481 gagtaattat attccagaaa cgccacctcc tggatatatc agtgaagatg gagaaacaag 3541 tgaccaacag ttgaatcaaa gtatggacac aggctctcca gcagaactat ctcctactac 3601 tctttcccct gttaatcata gcttggattt acagccagtt acttactcag aacctgcatt 3661 ttggtgttcg atagcatatt atgaattaaa tcagagggtt ggagaaacct tccatgcatc 3721 acagccctca ctcactgtag atggctttac agacccatca aattcagaga ggttctgctt 3781 aggtttactc tccaatgtta accgaaatgc cacggtagaa atgacaagaa ggcatatagg 3841 aagaggagtg cgcttatact acataggtgg ggaagttttt gctgagtgcc taagtgatag 3901 tgcaatcttt gtgcagagcc ccaattgtaa tcagagatat ggctggcacc ctgcaacagt 3961 gtgtaaaatt ccaccaggct gtaatctgaa gatcttcaac aaccaggaat ttgctgctct 4021 tctggctcag tctgttaatc agggttttga agccgtctat cagctaacta gaatgtgcac 4081 cataagaatg agttttgtga aagggtgggg agcagaatac cgaaggcaga cggtaacaag 4141 tactccttgc tggattgaac ttcatctgaa tggacctcta cagtggttgg acaaagtatt 4201 aactcagatg ggatcccctt cagtgcgttg ctcaagcatg tcaggatccg actacaaaga 4261 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtaatc 4321 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 4381 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 4441 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 4501 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 4561 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 4621 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 4681 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 4741 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 4801 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc 4861 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4921 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 4981 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5041 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5101 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5161 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5221 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5281 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5341 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5401 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt 5461 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5521 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5581 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5641 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5701 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5761 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5821 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 5881 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 5941 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6001 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6061 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 6121 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 6181 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6241 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc 6301 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca 6361 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 6421 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc 6481 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta 6541 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga 6601 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact 6661 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc 6721 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 6781 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 6841 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 6901 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 6961 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat 7021 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 7081 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 7141 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 7201 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 7261 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 7321 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 7381 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 7441 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 7501 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 7561 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 7621 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 7681 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 7741 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 7801 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 7861 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 7921 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 7981 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 8041 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 8101 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 8161 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 8221 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 8281 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 8341 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 8401 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 8461 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8521 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8581 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8641 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8701 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 8761 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 8821 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 8881 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 8941 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 9001 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 9061 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 9121 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 9181 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat 9241 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 9301 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat 9361 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta 9421 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata 9481 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact 9541 tagtagt //