LOCUS V028746 9886 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028746 VERSION V028746 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /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 1804..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..3912 /label="Bmpr1a(NM_009758)" /note="Bmpr1a(NM_009758)" /gene="Bmpr1a" CDS 3919..3984 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4028..4239 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4252..4520 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4549..4558 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4579..5241 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5311..5364 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5365..5964 /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 5965..6553 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6627..6860 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6932..7053 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7093..7228 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7261..7277) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7285..7301 /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(7309..7339) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7354..7375 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7663..8251) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8422..9282) /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(9283..9387) /gene="bla" /label="AmpR promoter" primer_bind 9861..9877 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga 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 ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatga ctcagctata cacttacatc 2341 agattactgg gagcctgtct gttcatcatt tctcatgttc aagggcagaa tctagatagt 2401 atgctccatg gcactggtat gaaatcagac ttggaccaga agaagccaga aaatggagtg 2461 actttagcac cagaggatac cttgcctttc ttaaagtgct attgctcagg acactgccca 2521 gatgatgcta ttaataacac atgcataact aatggccatt gctttgccat tatagaagaa 2581 gatgatcagg gagaaaccac attaacttct gggtgtatga agtatgaagg ctctgatttt 2641 caatgcaagg attcaccgaa agcccagcta cgcaggacaa tagaatgttg tcggaccaat 2701 ttgtgcaacc agtatttgca gcctacactg ccccctgttg ttataggtcc gttctttgat 2761 ggcagcatcc gatggctggt tgtgctcatt tccatggctg tctgtatagt tgctatgatc 2821 atcttctcca gctgcttttg ctataagcat tattgtaaga gtatctcaag caggggtcgt 2881 tacaaccgtg atttggaaca ggatgaagca tttattccag taggagaatc attgaaagac 2941 ctgattgacc agtcccaaag ctctgggagt ggatctggat tgcctttatt ggttcagcga 3001 actattgcca aacagattca gatggttcgg caggttggta aaggccgcta tggagaagta 3061 tggatgggta aatggcgtgg tgaaaaagtg gctgtcaaag tgttttttac cactgaagaa 3121 gctagctggt ttagagaaac agaaatctac cagacggtgt taatgcgtca tgaaaatata 3181 cttggtttta tagctgcaga cattaaaggc actggttcct ggactcagct gtatttgatt 3241 actgattacc atgaaaatgg atctctctat gacttcctga aatgtgccac actagacacc 3301 agagccctac tcaagttagc ttattctgct gcttgtggtc tgtgccacct ccacacagaa 3361 atttatggta cccaagggaa gcctgcaatt gctcatcgag acctgaagag caaaaacatc 3421 cttattaaga aaaatggaag ttgctgtatt gctgacctgg gcctagctgt taaattcaac 3481 agtgatacaa atgaagttga catacccttg aataccaggg tgggcaccaa gcggtacatg 3541 gctccagaag tgctggatga aagcctgaat aaaaaccatt tccagcccta catcatggct 3601 gacatctata gctttggttt gatcatttgg gaaatggctc gtcgttgtat tacaggagga 3661 atcgtggagg aatatcaatt accatattac aacatggtgc ccagtgaccc atcctatgag 3721 gacatgcgtg aggttgtgtg tgtgaaacgc ttgcggccaa tcgtgtctaa ccgctggaac 3781 agcgatgaat gtcttcgagc agttttgaag ctaatgtcag aatgttgggc ccataatcca 3841 gcctccagac tcacagcttt gagaatcaag aagacacttg caaaaatggt tgaatcccag 3901 gatgtaaaga ttggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3961 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 4021 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 4081 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 4141 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 4201 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 4261 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 4321 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4381 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4441 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4501 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4561 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4621 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4681 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4741 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4801 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4861 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4921 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4981 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 5041 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 5101 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 5161 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 5221 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 5281 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 5341 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 5401 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5461 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5521 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5581 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5641 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5701 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5761 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5821 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5881 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5941 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 6001 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 6061 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 6121 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 6181 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 6241 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 6301 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 6361 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6421 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6481 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6541 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6601 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6661 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6721 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6781 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6841 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6901 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6961 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 7021 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 7081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 7141 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 7201 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 7261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7441 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 8041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 8101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 8161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 8221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 8281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8341 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8401 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8461 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8521 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8581 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8641 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8701 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8761 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8821 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8881 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8941 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 9001 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 9061 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 9121 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 9181 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 9241 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9301 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9361 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9421 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9481 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9541 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9601 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9661 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9721 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9781 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9841 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //