LOCUS V024482 8896 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024482 VERSION V024482 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2815..3366 /label="FDX1(NM_004109)" /note="FDX1(NM_004109)" /gene="FDX1" CDS 3373..3438 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3470..4056 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 4057..4851 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 4865..5453 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5660..6293 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6421..6437) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6445..6461 /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(6469..6499) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6514..6535 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6823..7411) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7582..8442) /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(8443..8547) /gene="bla" /label="AmpR promoter" polyA_signal 8595..8729 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggct 2821 gccgctgggg gcgcccggct gctgcgcgcc gcttctgctg tcctcggcgg cccggccggc 2881 cggtggctgc accacgctgg gtcccgcgct ggatccagcg gcctgctgag gaaccggggg 2941 ccgggcggga gcgcggaggc gagccggtcg ctgagcgtgt cggcgcgggc ccggagcagc 3001 tcagaagata aaataacagt ccactttata aaccgtgatg gtgaaacatt aacaaccaaa 3061 ggaaaagttg gtgattctct gctagatgtt gtggttgaaa ataatctaga tattgatggc 3121 tttggtgcat gtgagggaac cctggcttgt tcaacctgtc acctcatctt tgaagatcac 3181 atatatgaga agttagatgc aatcactgat gaggagaatg acatgctcga tctggcatat 3241 ggactaacag acagatcacg gttgggctgc caaatctgtt tgacaaaatc tatggacaat 3301 atgactgttc gagtgcctga aacagtggct gatgccagac aatccattga tgtgggcaag 3361 acctccctcg aggactacaa agaccatgac ggtgattata aagatcatga catcgactac 3421 aaggatgacg atgacaagta gactagttct agagcggccg cggatcccgc ccctctccct 3481 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct 3541 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc 3601 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc 3661 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg 3721 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa 3781 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt 3841 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg 3901 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta 3961 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt 4021 tcctttgaaa aacacgatga taagcttgcc acaaccatgg ctgaacaaga tggattgcac 4081 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 4141 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 4201 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 4261 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4321 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4381 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 4441 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4501 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 4561 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 4621 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 4681 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 4741 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 4801 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg aacgcgtctg 4861 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4921 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4981 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5041 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5101 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5161 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5221 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct 5281 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5341 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5401 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat 5461 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct 5521 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc 5581 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5641 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt 5701 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg 5761 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca 5821 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg 5881 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca 5941 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct 6001 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg 6061 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct 6121 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 6181 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 6241 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 6301 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 6361 gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg cgtaatcatg 6421 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6481 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6541 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6601 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6661 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6721 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6781 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6841 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6901 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6961 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7021 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7081 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7141 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7201 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7261 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7321 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7381 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7441 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7501 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7561 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7621 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7681 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7741 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7801 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7861 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7921 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7981 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8041 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8101 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8161 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8221 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8281 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8341 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8401 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8461 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8521 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 8581 cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 8641 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 8701 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc 8761 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct 8821 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta 8881 ctacaaactt agtagt //