LOCUS V035965 8710 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035965 VERSION V035965 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3564 /label="Bmp10(NM_009756)" /note="Bmp10(NM_009756)" /gene="Bmp10" CDS 3571..3636 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3680..3891 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3904..4172 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4199..4798 /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 4805..5393 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5467..5700 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5772..5893 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5912..6047 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6085..6101) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6109..6125 /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(6133..6163) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6178..6199 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6487..7075) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7246..8106) /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(8107..8211) /gene="bla" /label="AmpR promoter" primer_bind 8685..8701 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggggtct ctggttctgc cgctgagcgc cgtcttctgc 2341 ctggtggctc actcggcttc tggcagcccc attatgggcc ttgagcagtc gcccctggaa 2401 gaagacatgc ccttctttga tgatatcttc acggagcaag atggtattga cttcaacaca 2461 ctgctgcaga gcatgaagaa cgagtttctc aagacgctga acttgtcgga cattcctgtg 2521 caggacacgg gcagagtgga tccaccagag tacatgctgg agctctacaa caaattcgcc 2581 acagaccgga cctccatgcc gtctgctaac atcatccgga gcttcaagaa cgaagatctg 2641 ttttctcaac cagtcacttt taatgggctc cggaaatatc ctctcctctt caatgtgtct 2701 atccctcacc atgaagaggt cgtcatggct gaactgcggt tgtacacgct ggtgcagaga 2761 gatcgcatga tgtatgatgg cgtggaccgt aaaattacca tttttgaggt actagagagt 2821 gcagacggta gcgaggagga gaggagcatg ctggtcttag tatcaacaga gatctacgga 2881 accaacagtg agtgggagac atttgacgtc acagatgcca ccagacgttg gcaaaagtca 2941 ggcccatcaa cccatcagct ggagatccac atagaaagca gacaaaacca agctgaggac 3001 accggaaggg gacaactgga aatagatatg agtgcccaaa ataagcatga ccctttgctg 3061 gttgtgtttt ctgatgacca aagcaatgac aaggagcaga aagaagaact gaacgaattg 3121 atcacccatg agcaggatct ggacctggac tcagatgctt tcttcagtgg gcccgatgaa 3181 gaggctctgc tgcagatgag gtcgaacatg attgatgatt cgtccgctcg gatcaggagg 3241 aacgccaagg ggaactactg taagaagacc ccactataca tcgacttcaa ggagattggg 3301 tgggactcct ggatcatcgc tcctcctggg tatgaagcct atgagtgccg gggtgtgtgt 3361 aactaccctc tggcggagca cctcacacct acaaaacacg caattattca ggccttggtc 3421 cacctcaaga attcccagaa agcttccaaa gcctgctgtg tgcccacgaa gctggatccc 3481 atctccatcc tctatttaga taaaggtgtc gtcacctaca agtttaaata tgaagggatg 3541 gctgtgtctg agtgtggctg tagaggatcc gactacaaag accatgacgg tgattataaa 3601 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3661 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3721 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3781 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3841 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3901 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3961 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4021 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4081 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4141 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat 4201 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 4261 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 4321 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4381 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4441 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4501 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4561 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4621 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4681 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4741 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 4801 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4861 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4921 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4981 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5041 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5101 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5161 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5221 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5281 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5341 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 5401 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5461 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5521 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5581 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5641 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5701 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5761 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5821 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5881 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 5941 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6001 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6061 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6121 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6181 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6241 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6901 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 6961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7201 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7261 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7321 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7381 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7441 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7501 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7561 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7621 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7681 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7741 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7801 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7861 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 7921 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7981 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8041 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8101 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8161 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8221 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8281 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8341 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8401 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8461 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8521 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8581 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8641 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8701 tgccaagctg //