LOCUS V039208 8884 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039208 VERSION V039208 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 1960..2171 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2184..2452 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 2492..2501 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2501..2566 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2582..4291 /label="SLC47A1(NM_018242)" /note="SLC47A1(NM_018242)" /gene="SLC47A1" CDS 4307..4360 /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 4367..4966 /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 4979..5567 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5641..5874 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5946..6067 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6086..6221 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6259..6275) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6283..6299 /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(6307..6337) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6352..6373 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6661..7249) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7420..8280) /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(8281..8385) /gene="bla" /label="AmpR promoter" primer_bind 8859..8875 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca 1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc 2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta 2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct 2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc 2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc 2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt 2461 gaccggcgcc tactctagag ctagcgaatt cgccaccatg gactacaaag accatgacgg 2521 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagcttg gtaccctcga 2581 gatggaagct cctgaggagc ccgcgccagt gcgcggaggc ccggaggcca cccttgaggt 2641 ccgtgggtcg cgctgcttgc ggctgtccgc cttccgagaa gagctgcggg cgctcttggt 2701 cctggctggc cccgcgttct tggttcagct gatggtgttc ctgatcagct tcataagctc 2761 cgtgttctgt ggccacctgg gcaagctgga gctggatgca gtcacgctgg caatcgcggt 2821 tatcaatgtc actggtgtct cagtgggatt cggcttatct tctgcctgtg acaccctcat 2881 ctcccagacg tacgggagcc agaacctgaa gcacgtgggc gtgatcctgc agcggagtgc 2941 gctcgtcctg ctcctctgct gcttcccctg ctgggcgctc tttctcaaca cccagcacat 3001 cctgctgctc ttcaggcagg acccagatgt gtccaggctt acccagacct atgtcacgat 3061 cttcattcca gctcttcctg caacctttct ttatatgtta caagttaaat atttgctcaa 3121 ccagggaatt gtactgcccc agatcgtaac tggagttgca gccaaccttg tcaatgccct 3181 cgccaactat ctgtttctcc atcaactgca tcttggggtg ataggctctg cactggcaaa 3241 cttgatttcc cagtacaccc tggctctact cctctttctc tacatcctcg ggaaaaaact 3301 gcatcaagct acatggggag gctggtccct cgagtgcctg caggactggg cctccttcct 3361 ccgcctggcc atccccagca tgctcatgct gtgcatggag tggtgggcct atgaggtcgg 3421 gagcttcctc agtggcatcc tcggcatggt ggagctgggc gctcagtcca tcgtgtatga 3481 actggccatc attgtgtaca tggtccctgc aggcttcagt gtggctgcca gtgtccgggt 3541 aggaaacgct ctgggtgctg gagacatgga gcaggcacgg aagtcctcta ccgtttccct 3601 gctgattaca gtgctctttg ctgtagcctt cagtgtcctg ctgttaagct gtaaggatca 3661 cgtggggtac atttttacta ccgaccgaga catcattaat ctggtggctc aggtggttcc 3721 aatttatgct gtttcccacc tctttgaagc tcttgcttgc acgagtggtg gtgttctgag 3781 ggggagtgga aatcagaagg ttggagccat tgtgaatacc attgggtact atgtggttgg 3841 cctccccatc gggatcgcgc tgatgtttgc aaccacactt ggagtgatgg gtctgtggtc 3901 agggatcatc atctgtacag tctttcaagc tgtgtgtttt ctaggcttta ttattcagct 3961 aaattggaaa aaagcctgtc agcaggctca ggtacacgcc aatttgaaag taaacaacgt 4021 gcctcggagt gggaattctg ctctccctca ggatccgctt cacccagggt gccctgaaaa 4081 ccttgaagga attttaacga acgatgttgg aaagacaggc gagcctcagt cagatcagca 4141 gatgcgccaa gaagaacctt tgccggaaca tccacaggac ggcgctaaat tgtccaggaa 4201 acagctggtg ctgcggcgag ggcttctgct cctgggggtc ttcttaatct tgctggtggg 4261 gattttagtg agattctatg tcagaattca gggatccgcg gccgctgagg gcagaggaag 4321 tcttctaaca tgcggtgacg tggaggagaa tcccggccct tccgggatga ccgagtacaa 4381 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc 4441 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg 4501 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg 4561 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg 4621 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc 4681 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct 4741 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct 4801 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc 4861 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc 4921 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaatct aggtcgacaa 4981 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5041 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5101 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5161 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5221 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5281 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5341 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5401 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5461 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5521 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5581 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5641 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5701 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5761 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5821 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5881 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 5941 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6001 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6061 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6121 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6181 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6241 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6301 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6361 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6421 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6481 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6841 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7081 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8401 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8461 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8521 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8581 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8641 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8701 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8761 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8821 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8881 gctg //