LOCUS V037908 8924 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037908 VERSION V037908 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" regulatory 2288..2297 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2297..2362 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2369..3826 /label="Slc16a12(NM_172838)" /note="Slc16a12(NM_172838)" /gene="Slc16a12" promoter 3894..4105 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4118..4386 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4413..5012 /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 5019..5607 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5681..5914 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5986..6107 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6126..6261 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6299..6315) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6323..6339 /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(6347..6377) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6392..6413 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6701..7289) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7460..8320) /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(8321..8425) /gene="bla" /label="AmpR promoter" primer_bind 8899..8915 /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga 2341 ctacaaggat gacgatgaca aggaattcat gacgaagata actagagtca gcttggcatc 2401 ccctccagat ggaggctggg gctggatgat agtggccggc tgcttccttg ttaccatctg 2461 cacccgggca gtcaccagat gtatctccat attttttgtg gagtttcaga catactttgc 2521 tcaggattac tcccaaacag catggatcca ttccattgta gactgtatga caatgctctg 2581 tgctccgctt gggagtgttg tcagtaacca gttatcctgt caagcgggga tcatgctggg 2641 tggcttgctg gcatctactg gtttcatcct gggctcattt gccaccagct tgaagcacct 2701 ctacctcagc ctgggagttc ttacaggtct tggatttgca ctttgttact ctccggcaat 2761 tgccatggtg ggcaagtatt tcagcagacg gaaagccctg gcttatggga ttgccatgtc 2821 aggcagtggc attggtacct tcatcctggc tcctgtcgtt cagctcctca tcgaacagtt 2881 ttcctggcga ggagcactgc tcattcttgg cgggttcgtt ctgaatctct gcgtctgcgg 2941 cgctctgatg cgtccaatta cccttaaaga ggaccgatca gtcccagaaa agaatcacaa 3001 ccgtgaatct cagcgagaag actgtaagca agcatctccc tattcacctt tgaccaaaga 3061 atgtacagag acccgtcttt gctgctcctt gcagcaggaa tacggttttt tactgatgtc 3121 agacttcgtc gtgttggccg tctctgttct gttcatggcc tacggctgca gccctctctt 3181 tgtgtaccta gtgccttatg ctttgagtgt cggagtgagc caccatcaag ctgctttcct 3241 catgtccatc ctcggggtaa tcgatattgt tggcaatatc acatttggct ggctaactga 3301 cagaaggtgt ctgaagaatt accgatatgt ttgctacctc tttgccgtgg cactagatgg 3361 gctgtgctat ctgtgtcttc caatgcttca aaccttcccc cttcttgtgc ctttctcttg 3421 tacctttggc tactttgatg gagcctacgt taccttgatc ccggtcgtga ctgcagagat 3481 agtagggacc acctctttgt cttcagcact tggcgtggtc tacttccttc atgcagtgcc 3541 atacttggtg agcccaccca ttgcaggatg gctggtggac acgacgggca gctacactgc 3601 agccttcctt ctctgtggat ttgccatgat atttagttcc atcctgcttg gcttcgttag 3661 aattgtcaag aggatgaaga gaacacaagt gcctttcccg gtcaaagact cagatcccaa 3721 gctacaactg tggaccaatg gatcagtggc ctactctgta gcaagggagt tagatcagaa 3781 agatgaggaa cccctgccta aagccaggtc tggttgcaac ctcacgtgag gatccctcga 3841 gagatcagtg tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga 3901 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc 3961 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt 4021 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg 4081 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg 4141 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc 4201 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag 4261 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct 4321 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta 4381 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc 4441 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact 4501 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc 4561 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 4621 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 4681 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 4741 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 4801 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 4861 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 4921 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 4981 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa 5041 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 5101 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 5161 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 5221 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 5281 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 5341 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 5401 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc 5461 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 5521 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 5581 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 5641 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5701 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 5761 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5821 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5881 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 5941 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6001 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6061 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6121 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6181 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6241 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7141 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7561 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8161 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8221 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8281 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8341 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8401 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8461 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8521 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8581 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8641 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8701 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 8761 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 8821 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 8881 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //