LOCUS V043158 8804 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043158 VERSION V043158 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..3636 /label="TUBB4B(NM_006088)" /note="TUBB4B(NM_006088)" /gene="TUBB4B" CDS 3643..3669 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3673..3699 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3703..3729 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3774..3985 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3998..4266 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4293..4892 /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 4899..5487 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5561..5794 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5866..5987 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6006..6141 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6179..6195) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6203..6219 /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(6227..6257) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6272..6293 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6581..7169) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7340..8200) /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(8201..8305) /gene="bla" /label="AmpR promoter" primer_bind 8779..8795 /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 catgagggaa atcgtgcact tgcaggccgg gcagtgcggc 2341 aaccaaatcg gcgccaagtt ttgggaggtg atcagcgatg agcacggcat cgaccccacg 2401 ggcacctacc acggggacag cgacctgcag ctggaacgca tcaacgtgta ctacaatgag 2461 gccaccggcg gcaagtacgt gccccgcgcc gtgctcgtgg atctggagcc cggcaccatg 2521 gactccgtgc gctcggggcc cttcgggcag atcttccggc cggacaactt cgttttcggt 2581 cagagtggtg ctgggaacaa ctgggccaag gggcactaca cagaaggcgc ggagctggtg 2641 gactcggtgc tggatgttgt gagaaaggag gctgagagct gtgactgcct gcagggtttc 2701 cagctgaccc actccctggg tggggggact gggtctggga tgggtaccct cctcatcagc 2761 aagatccggg aggagtaccc agacaggatc atgaacacgt ttagtgtggt gccttcgccc 2821 aaagtgtcag acacagtggt ggagccctac aacgccaccc tctcagtcca ccagctcgta 2881 gaaaacacag acgagaccta ctgcattgat aacgaagctc tctacgacat ttgcttcaga 2941 accctaaagc tgaccacgcc cacctatggt gacctgaacc acctggtgtc tgctaccatg 3001 agtggggtca ccacctgcct gcgcttccca ggccagctca atgctgacct gcggaagctg 3061 gctgtgaaca tggtcccgtt tccccggctg cacttcttca tgcccggctt tgccccactg 3121 accagccggg gcagccagca gtaccgggcg ctgaccgtgc ccgagctcac ccagcagatg 3181 tttgatgcca agaacatgat ggctgcctgc gacccccgcc atggccgcta cctgacggtt 3241 gccgccgtgt tcaggggccg catgtccatg aaggaggtgg atgagcaaat gcttaatgtc 3301 caaaacaaaa acagcagcta ttttgttgag tggatcccca acaatgtgaa aacggctgtc 3361 tgtgacatcc cacctcgggg gctaaaaatg tccgccacct tcattggcaa cagcacggcc 3421 atccaggagc tgttcaagcg catctccgag cagttcacgg ccatgttccg gcgcaaggcc 3481 ttcctgcact ggtacacggg cgagggcatg gacgagatgg agttcaccga ggccgagagc 3541 aacatgaatg acctggtgtc cgagtaccag cagtaccagg atgccacagc cgaggaggag 3601 ggcgagttcg aggaggaggc tgaggaggag gtggccggat cctacccata cgacgtacct 3661 gattatgcaa cttatcctta tgatgtacct gattatgcta gctatcctta tgatgtccca 3721 gactacgcat aggcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga 3781 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc 3841 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt 3901 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg 3961 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg 4021 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc 4081 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag 4141 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct 4201 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta 4261 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc 4321 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact 4381 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc 4441 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 4501 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 4561 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 4621 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 4681 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 4741 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 4801 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 4861 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa 4921 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 4981 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 5041 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 5101 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 5161 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 5221 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 5281 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc 5341 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 5401 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 5461 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 5521 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5581 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 5641 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5701 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5761 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 5821 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 5881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 5941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6001 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6061 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6121 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6181 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6241 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6301 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6361 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7021 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7261 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7321 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7381 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7441 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7501 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7561 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7621 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7681 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7741 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7801 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7861 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7921 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7981 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8041 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8101 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8161 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8281 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8341 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8401 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8461 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8521 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8581 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 8641 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 8701 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 8761 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //