LOCUS V029973 9106 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029973 VERSION V029973 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..3960 /label="IL20RA(NM_014432)" /note="IL20RA(NM_014432)" /gene="IL20RA" CDS 3967..4032 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4076..4287 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4300..4568 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4595..5194 /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 5201..5789 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5863..6096 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6168..6289 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6308..6443 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6481..6497) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6505..6521 /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(6529..6559) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6574..6595 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6883..7471) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7642..8502) /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(8503..8607) /gene="bla" /label="AmpR promoter" primer_bind 9081..9097 /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 catgcgggct cccggccgcc cggccctgcg gccgctgccg 2341 ctgccgccgc tgctgctgtt gctcctggcg gcgccttggg gacgggcagt tccctgtgtc 2401 tctggtggtt tgcctaaacc tgcaaacatc accttcttat ccatcaacat gaagaatgtc 2461 ctacaatgga ctccaccaga gggtcttcaa ggagttaaag ttacttacac tgtgcagtat 2521 ttcatatatg ggcaaaagaa atggctgaat aaatcagaat gcagaaatat caatagaacc 2581 tactgtgatc tttctgctga aacttctgac tacgaacacc agtattatgc caaagttaag 2641 gccatttggg gaacaaagtg ttccaaatgg gctgaaagtg gacggttcta tcctttttta 2701 gaaacacaaa ttggcccacc agaggtggca ctgactacag atgagaagtc catttctgtt 2761 gtcctgacag ctccagagaa gtggaagaga aatccagaag accttcctgt ttccatgcaa 2821 caaatatact ccaatctgaa gtataacgtg tctgtgttga atactaaatc aaacagaacg 2881 tggtcccagt gtgtgaccaa ccacacgctg gtgctcacct ggctggagcc gaacactctt 2941 tactgcgtac acgtggagtc cttcgtccca gggccccctc gccgtgctca gccttctgag 3001 aagcagtgtg ccaggacttt gaaagatcaa tcatcagagt tcaaggctaa aatcatcttc 3061 tggtatgttt tgcccgtatc tattaccgtg tttctttttt ctgtgatggg ctattccatc 3121 taccgatata tccacgttgg caaagagaaa cacccagcaa atttgatttt gatttatgga 3181 aatgaatttg acaaaagatt ctttgtgcct gctgaaaaaa tcgtgattaa ctttatcacc 3241 ctcaatatct cggatgattc taaaatttct catcaggata tgagtttact gggaaaaagc 3301 agtgatgtat ccagccttaa tgatcctcag cccagcggga acctgaggcc ccctcaggag 3361 gaagaggagg tgaaacattt agggtatgct tcgcatttga tggaaatttt ttgtgactct 3421 gaagaaaaca cggaaggtac ttctctcacc cagcaagagt ccctcagcag aacaataccc 3481 ccggataaaa cagtcattga atatgaatat gatgtcagaa ccactgacat ttgtgcgggg 3541 cctgaagagc aggagctcag tttgcaggag gaggtgtcca cacaaggaac attattggag 3601 tcgcaggcag cgttggcagt cttgggcccg caaacgttac agtactcata cacccctcag 3661 ctccaagact tagaccccct ggcgcaggag cacacagact cggaggaggg gccggaggaa 3721 gagccatcga cgaccctggt cgactgggat ccccaaactg gcaggctgtg tattccttcg 3781 ctgtccagct tcgaccagga ttcagagggc tgcgagcctt ctgaggggga tgggctcgga 3841 gaggagggtc ttctatctag actctatgag gagccggctc cagacaggcc accaggagaa 3901 aatgaaacct atctcatgca attcatggag gaatgggggt tatatgtgca gatggaaaac 3961 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat 4021 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca 4081 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt 4141 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt 4201 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt 4261 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca 4321 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct 4381 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg 4441 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg 4501 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt 4561 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg 4621 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga 4681 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct 4741 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga 4801 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc 4861 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat 4921 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg 4981 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga 5041 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc 5101 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg 5161 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa 5221 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 5281 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 5341 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 5401 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 5461 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 5521 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 5581 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 5641 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 5701 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 5761 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg 5821 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 5881 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 5941 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 6001 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 6061 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 6121 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 6181 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 6241 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 6301 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 6361 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 6421 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg 6481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7321 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8641 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8701 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8761 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8821 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 8881 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 8941 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9001 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9061 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //