LOCUS V035917 7750 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035917 VERSION V035917 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..2799 /label="TXN2(NM_012473)" /note="TXN2(NM_012473)" /gene="TXN2" CDS 2806..2871 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2915..3126 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3139..3407 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3440..3838 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 3845..4433 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4507..4740 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4812..4933 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 4952..5087 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5125..5141) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5149..5165 /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(5173..5203) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5218..5239 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5527..6115) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6286..7146) /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(7147..7251) /gene="bla" /label="AmpR promoter" primer_bind 7725..7741 /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 catggctcag cgacttcttc tgaggaggtt cctggcctct 2341 gtcatctcca ggaagccctc tcagggtcag tggccacccc tcacttccag agccctgcag 2401 accccacaat gcagtcctgg tggcctgact gtaacaccca acccagcccg gacaatatac 2461 accacgagga tctccttgac aacctttaat atccaggatg gacctgactt tcaagaccga 2521 gtggtcaaca gtgagacacc agtggttgtg gatttccacg cacagtggtg tggaccctgc 2581 aagatcctgg ggccgaggtt agagaagatg gtggccaagc agcacgggaa ggtggtgatg 2641 gccaaggtgg atattgatga ccacacagac ctcgccattg agtatgaggt gtcagcggtg 2701 cccactgtgc tggccatgaa gaatggggac gtggtggaca agtttgtggg catcaaggat 2761 gaggatcagt tggaggcctt cctgaagaag ctgattggcg gatccgacta caaagaccat 2821 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 2881 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 2941 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3001 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3061 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3121 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3181 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3241 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3301 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3361 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 3421 gcgcctacgc taggccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga 3481 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc 3541 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg 3601 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac 3661 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg 3721 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca 3781 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt 3841 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 3901 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 3961 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4021 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 4081 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 4141 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 4201 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 4261 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 4321 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 4381 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 4441 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 4501 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 4561 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 4621 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 4681 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 4741 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 4801 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4861 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4921 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 4981 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 5041 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 5101 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 5161 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 5221 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 5281 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 5341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 5401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 5461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 5521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 5581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 5641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 5701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 5761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 5821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 5881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 5941 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 6001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 6061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 6121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 6181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 6241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 6301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 6361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 6421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 6481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 6541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 6601 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 6661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 6721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 6781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 6841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 6901 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 6961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 7081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 7141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 7201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 7261 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 7321 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 7381 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 7441 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 7501 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 7561 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 7621 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 7681 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 7741 tgccaagctg //