LOCUS V022657 8818 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022657 VERSION V022657 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..3684 /label="CORO1A(NM_007074)" /note="CORO1A(NM_007074)" /gene="CORO1A" CDS 3685..3708 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 3709..3738 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" promoter 3788..3999 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4012..4280 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4307..4906 /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 4913..5501 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5575..5808 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5880..6001 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6020..6155 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6193..6209) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6217..6233 /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(6241..6271) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6286..6307 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6595..7183) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7354..8214) /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(8215..8319) /gene="bla" /label="AmpR promoter" primer_bind 8793..8809 /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 catgagccgg caggtggtcc gctccagcaa gttccgccac 2341 gtgtttggac agccggccaa ggccgaccag tgctatgaag atgtgcgcgt ctcacagacc 2401 acctgggaca gtggcttctg tgctgtcaac cctaagtttg tggccctgat ctgtgaggcc 2461 agcgggggag gggccttcct ggtgctgccc ctgggcaaga ctggacgtgt ggacaagaat 2521 gcgcccacgg tctgtggcca cacagcccct gtgctagaca tcgcctggtg cccgcacaat 2581 gacaacgtca ttgccagtgg ctccgaggac tgcacagtca tggtgtggga gatcccagat 2641 gggggcctga tgctgcccct gcgggagccc gtcgtcaccc tggagggcca caccaagcgt 2701 gtgggcattg tggcctggca caccacagcc cagaacgtgc tgctcagtgc aggttgtgac 2761 aacgtgatca tggtgtggga cgtgggcact ggggcggcca tgctgacact gggcccagag 2821 gtgcacccag acacgatcta cagtgtggac tggagccgag atggaggcct catttgtacc 2881 tcctgccgtg acaagcgcgt gcgcatcatc gagccccgca aaggcactgt cgtagctgag 2941 aaggaccgtc cccacgaggg gacccggccc gtgcgtgcag tgttcgtgtc ggaggggaag 3001 atcctgacca cgggcttcag ccgcatgagt gagcggcagg tggcgctgtg ggacacaaag 3061 cacctggagg agccgctgtc cctgcaggag ctggacacca gcagcggtgt cctgctgccc 3121 ttctttgacc ctgacaccaa catcgtctac ctctgtggca agggtgacag ctcaatccgg 3181 tactttgaga tcacttccga ggcccctttc ctgcactatc tctccatgtt cagttccaag 3241 gagtcccagc ggggcatggg ctacatgccc aaacgtggcc tggaggtgaa caagtgtgag 3301 atcgccaggt tctacaagct gcacgagcgg aggtgtgagc ccattgccat gacagtgcct 3361 cgaaagtcgg acctgttcca ggaggacctg tacccaccca ccgcagggcc cgaccctgcc 3421 ctcacggctg aggagtggct ggggggtcgg gatgctgggc ccctcctcat ctccctcaag 3481 gatggctacg tacccccaaa gagccgggag ctgagggtca accggggcct ggacaccggg 3541 cgcaggaggg cagcaccaga ggccagtggc actcccagct cggatgccgt gtctcggctg 3601 gaggaggaga tgcggaagct ccaggccacg gtgcaggagc tccagaagcg cttggacagg 3661 ctggaggaga cagtccaggc caaggattac aaggatgacg acgataagga acaaaaactc 3721 atctcagaag aggatctgta ggatccgcgg ccgcgaagga tctgcgatcg ctccggtgcc 3781 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3841 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3901 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3961 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 4021 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 4081 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4141 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4201 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4261 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa 4321 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc 4381 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg 4441 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg 4501 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg 4561 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc 4621 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct 4681 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct 4741 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc 4801 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc 4861 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc 4921 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 4981 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 5041 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 5101 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 5161 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 5221 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 5281 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt 5341 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 5401 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 5461 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa 5521 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 5581 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt 5641 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 5701 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 5761 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 5821 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 5881 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 5941 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 6001 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc 6061 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 6121 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa 6181 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6241 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6301 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6361 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6421 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6481 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6541 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6601 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6661 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6721 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6781 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6841 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6901 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6961 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7021 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7081 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 7141 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7201 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7261 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7321 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7381 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7441 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7501 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7561 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7621 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7681 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7741 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7801 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7861 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7921 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 7981 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8041 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 8101 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 8161 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8221 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8281 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8341 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8401 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8461 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8521 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8581 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8641 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8701 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 8761 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //