LOCUS V021427 10104 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021427 VERSION V021427 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 2301..5006 /label="Aldh1l1(NM_027406)" /note="Aldh1l1(NM_027406)" /gene="Aldh1l1" CDS 5007..5030 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 5074..5285 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5298..5566 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5593..6192 /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 6199..6787 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6861..7094 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7166..7287 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7306..7441 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7479..7495) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7503..7519 /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(7527..7557) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7572..7593 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7881..8469) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8640..9500) /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(9501..9605) /gene="bla" /label="AmpR promoter" primer_bind 10079..10095 /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta 2281 gagctagcga attcgccacc atgaagattg cagtaatcgg gcagagcctg tttggccagg 2341 aggtttactg ccagctaagg aaggagggcc atgaggtggt gggtgtgttc accatcccag 2401 acaaggatgg gaaagcagat cccctgggcc tggaggccga gaaggatggc gtgcccgtgt 2461 ttaagttccc tcggtggcgg gctcgaggac aggctctgcc cgaggtcgtg gccaagtacc 2521 aggctctggg tgcagaactc aacgtgctcc ccttctgcag ccagttcata cccatggagg 2581 tcatcaatgc cccccggcat ggctccatca tctaccatcc atctctgctg cccaggcacc 2641 gaggggcctc agccatcaac tggaccctca ttcatggaga taagaaagga ggcttcacta 2701 tcttctgggc tgatgacggt ctggacactg gtgaccttct gctgcagaag gagtgtgacg 2761 tgcttccaga tgacaccgtg agcacgctgt acaaccgttt cctcttccca gagggcatca 2821 aagggatggt tcaggctgtg cgactgattg cagagggcac agccccacgc cgtccccagc 2881 ctgaggaagg agccacctat gagggcattc agaaaaagga gacagccatg atcaactggg 2941 accagccagc agaggccatt cacaactgga tccgtgggaa tgacaaggtg ccaggtgcct 3001 ggacagaggc ttgtgggcag aagctgacgt ttttcaactc gacactcaac acttcaggac 3061 tggtggctca gggagaagct ctacccatcc caggagccca tcgtccaggc ctagtcacca 3121 aagcgggact catcctcttt gggaatgatg atagaatgtt gctggtgaag aatatccagc 3181 tggaagatgg caagatgatg ccagcctccc agttcttcaa ggggtctgct agcagtgccc 3241 tggagctgac cgaggaggag ctggccacag cagaggccgt gcggagctct tggatgcgaa 3301 ttctgcccaa tgtcccagag gtagaagact ctacagattt cttcaagtca ggagctgcat 3361 ccgtagatgt tgtgaggctg gtggaggagg tgaaggagct gtgtgacggg ctggagttag 3421 aaaatgagga tgtttacatg gccaccacct tcggggactt catccagctc ctagtgagga 3481 agttgagagg ggaggacggc gagagcgagt gtgtcattaa ctacgtggaa aaggcagtga 3541 agaaactgac tctccaaatg ccctaccagc tcttcatagg cggcgagttt gtggatgctg 3601 agggcgcgaa gacctacagt accataaacc caacggatgg aagtgtcatc tgccaggtgt 3661 ctctagctca ggtgagtgat gttgacaagg cggtggcagc agcgaaggag gcctttgaga 3721 atggactgtg gggaaagata aatgcgcgtg accggggccg gctcctgtac aggttggcgg 3781 acctcatgga gcagcaccag gaagagctag ccaccattga ggccctggac gcaggtgccg 3841 tctacacgct ggccctgaag acgcatgtgg gcatgtccat ccagaccttc cgatacttcg 3901 ctggctggtg tgataagatc cagggtgcca ccatccccat caaccaggct agacccaacc 3961 gcaacctgac cttgaccaag aaggaacctg ttggggtctg tggtattgtc atcccctgga 4021 actatccctt aatgatgctg tcctggaaga cagcagcctg cctggctgcc gggaacaccg 4081 tggtgatcaa gcctgcccag gtgaccccac tcacagcctt gaagtttgca gagctgacac 4141 tgaaggctgg cattcccaag ggtgtggtca acatcctccc aggatctggc tcgctggttg 4201 gccagagact ctcagaccac cctgatgtga ggaaaatagg gttcacaggc tccacggagg 4261 tgggaaaaca catcatgaaa agctgtgccc tgagtaatgt gaagaaggtc tccctggagc 4321 tgggtggaaa gtcacccctt atcatctttg ctgactgtga cctcaacaaa gctgtgcaga 4381 tgggcatgag ctccgttttc ttcaacaaag gggagaactg cattgcggca ggccggctct 4441 ttgtggagga ctccatccat gaccagtttg tgcagaaagt ggtggaggaa gtagggaaga 4501 tgaaaatcgg caaccccctg gacagagata ccaaccatgg cccgcagaac catgaggccc 4561 acctgaggaa gctggtggag tattgccaac gtggtgtgaa ggaaggggcc acactggtct 4621 gtggtgggaa ccaagtccca aggccaggct tcttctttca gccaaccgtt ttcacagacg 4681 tggaggacca catgtacatc gctaaggagg agtccttcgg gcccatcatg atcatctctc 4741 ggtttgctga tggggacgtg gatgcagtgc tatctcgggc caatgctaca gaatttggcc 4801 tggcctctgg tgtcttcact cgggatatca acaaggccct gtatgtcagt gacaaactgc 4861 aggcgggcac tgtgtttgtc aacacataca acaagaccga tgtggccgca ccttttggag 4921 gattcaagca gtctggattt ggcaaagacc tgggagaggc ggccctgaat gagtacctgc 4981 ggatcaagac tgtgactttt gagtacgatt acaaggacga cgatgacaag tagcggccgc 5041 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc 5101 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg 5161 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac 5221 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa 5281 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 5341 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 5401 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 5461 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 5521 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg 5581 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 5641 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 5701 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 5761 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 5821 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 5881 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac 5941 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 6001 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg 6061 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 6121 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 6181 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 6241 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 6301 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 6361 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 6421 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 6481 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 6541 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 6601 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 6661 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 6721 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 6781 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 6841 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 6901 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6961 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 7021 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 7081 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 7141 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 7201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 7261 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 7321 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 7381 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 7441 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 7501 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 7561 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 7621 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 7681 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 7741 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 7801 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 7861 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 7921 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 7981 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 8041 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 8101 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 8161 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 8221 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 8281 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 8341 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 8401 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 8461 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 8521 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 8581 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 8641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 8701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 8761 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 8821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 8881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 8941 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 9001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 9061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 9121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 9181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 9241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 9301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 9361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 9421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 9481 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 9541 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 9601 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 9661 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 9721 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 9781 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 9841 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 9901 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 9961 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 10021 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 10081 aaaacgacgg ccagtgccaa gctg //