LOCUS V032757 10094 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032757 VERSION V032757 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 2287..2352 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2377..5019 /label="EPB41L1(NM_012156)" /note="EPB41L1(NM_012156)" /gene="EPB41L1" promoter 5064..5275 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5288..5556 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5583..6182 /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 6189..6777 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6851..7084 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7156..7277 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7296..7431 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7469..7485) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7493..7509 /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(7517..7547) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7562..7583 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7871..8459) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8630..9490) /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(9491..9595) /gene="bla" /label="AmpR promoter" primer_bind 10069..10085 /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 agaatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat 2341 gacgatgaca agtctagaga attcggatcc gccaccatga caacagagac aggccccgac 2401 tctgaggtga agaaagctca ggaggaggcc ccgcagcagc ccgaggctgc tgccgctgtg 2461 accacccctg tgacccctgc aggccacggc cacccagagg ccaactccaa tgagaagcat 2521 ccatcccagc aggacacgcg gcctgctgaa cagagcctag acatggagga gaaggactac 2581 agtgaggccg atggcctttc ggagaggacc acgcccagca aggcccagaa atcgccccag 2641 aagattgcca agaaatacaa gagtgccatc tgccgggtca ctctgcttga tgcctcggag 2701 tatgagtgtg aggtggagaa acatggccgg ggccaggtgc tgtttgacct ggtctgtgaa 2761 cacctcaacc tcctagagaa ggactacttc ggcctgacct tctgtgatgc tgacagccag 2821 aagaactggc tggacccctc caaggagatc aagaagcaga tccggagtag cccctggaat 2881 tttgccttca cagtcaagtt ctacccgcct gatcctgccc agctgacaga agacatcaca 2941 agatactacc tgtgcctgca gctgcgggca gacatcatca cgggccggct gccatgctcc 3001 tttgtcacgc atgccctact gggctcctac gctgtgcagg ctgagctggg tgactatgat 3061 gctgaggagc atgtgggcaa ctatgtcagc gagctccgct tcgcccctaa ccagacccgg 3121 gagctggagg agaggatcat ggagctgcat aagacatata gggggatgac cccgggagaa 3181 gcagaaatcc acttcttaga gaatgccaag aagctttcca tgtacggagt agacctgcac 3241 catgccaagg actctgaggg catcgacatc atgttaggcg tttgtgccaa tggcctgctc 3301 atctaccggg accggctgag aatcaaccgc tttgcctggc ccaagatcct caagatctcc 3361 tacaagagga gtaacttcta tatcaagatc cggcctgggg agtatgagca atttgagagc 3421 acaattggct ttaagctccc aaaccaccgg tcagccaaga gactgtggaa ggtctgcatc 3481 gagcatcata cattcttccg gctggtgtcc cctgagcccc cacccaaggg cttcctggtg 3541 atgggctcca agttccggta cagtgggagg acccaggcac agactcgcca ggccagcgcc 3601 ctcattgacc ggcctgcacc cttctttgag cgttcttcca gcaaacggta caccatgtcc 3661 cgcagccttg atggagcaga gttctcccgc ccagcctcgg tcagcgagaa ccatgatgca 3721 gggcctgacg gtgacaagcg ggatgaggat ggcgagtctg gggggcaacg gtcagaggct 3781 gaggagggag aggtcaggac tccaaccaag atcaaggagc taaagccgga gcaggaaacc 3841 acgccgagac acaagcagga gttcttagac aagccagaag atgtcttgct gaagcaccag 3901 gccagcatca atgagctcaa aaggaccctg aaggagccca acagcaaact catccaccgg 3961 gatcgagact gggaacggga gcgcaggctg ccctcctccc ccgcctcccc ctcccccaag 4021 ggcacccctg agaaagccaa tgagagagca gggctgaggg agggctccga ggagaaagtc 4081 aaaccaccac gtccccgggc cccagagagt gacacaggcg atgaggacca ggaccaggag 4141 agggacacgg tgttcctgaa ggacaaccac ctggccattg agcgcaagtg ctccagcatc 4201 acggtcagct ctacgtctag cctggaggct gaggtggact tcacggtcat tggtgactac 4261 catggcagcg ccttcgaaga cttctcccgc agcctgcctg agctcgaccg ggacaaaagc 4321 gactcggaca ctgagggcct gctgttctcc cgggatctca acaagggggc ccccagccag 4381 gatgatgagt ctgggggcat tgaggacagc ccggatcgag gggcctgctc caccccggat 4441 atgccccagt ttgagcccgt gaaaacagaa accatgactg tcagcagtct ggccattaga 4501 aagaagattg agccggaggc cgtactgcag accagagtct ccgctatgga taacacccag 4561 caggttgatg ggagtgcctc agtggggagg gagttcatag caaccactcc ctccatcacc 4621 acggagacca tatcgaccac catggagaac agtctcaagt ccgggaaggg ggcagctgcc 4681 atgatcccag gcccacagac ggtggccacg gaaatccgtt ctctttctcc gatcatcggg 4741 aaagatgtcc tcaccagcac ctacggcgcc actgcggaaa ccctctcaac ctccaccacc 4801 acccatgtca ccaaaactgt gaaaggaggg ttttctgaga caaggatcga gaagcgaatc 4861 atcattactg gggatgaaga tgtcgatcaa gaccaggccc tggctttggc catcaaggag 4921 gccaaactgc agcatcctga tatgctggta accaaagctg tcgtatacag agaaacagac 4981 ccatccccag aggagaggga caagaagcca caggaatcct aagcggccgc gaaggatctg 5041 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt 5101 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga 5161 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag 5221 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa 5281 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc 5341 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc 5401 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag 5461 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt 5521 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct 5581 agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg 5641 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg 5701 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 5761 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 5821 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 5881 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 5941 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 6001 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 6061 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 6121 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 6181 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 6241 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 6301 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 6361 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 6421 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 6481 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 6541 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 6601 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 6661 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 6721 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 6781 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 6841 aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt 6901 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 6961 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 7021 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 7081 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata 7141 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc 7201 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 7261 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt 7321 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 7381 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt 7441 gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc 7501 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 7561 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 7621 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 7681 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 7741 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 7801 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 7861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 7921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 7981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 8041 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 8101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 8161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 8221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 8281 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 8341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 8401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 8461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 8521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 8581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 8641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 8701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 8761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 8821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 8881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 8941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 9001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 9061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 9121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 9181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 9241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 9301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 9361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 9421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 9481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 9541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 9601 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 9661 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc 9721 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 9781 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg 9841 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat 9901 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg 9961 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 10021 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 10081 ccagtgccaa gctg //