LOCUS V043481 9526 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043481 VERSION V043481 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..3552 /label="Ikbkg(NM_199103)" /note="Ikbkg(NM_199103)" /gene="Ikbkg" CDS 3559..3624 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3668..3879 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3892..4160 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4189..4198 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4219..4881 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 4951..5004 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5005..5604 /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 5605..6193 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6267..6500 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6572..6693 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6733..6868 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6901..6917) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6925..6941 /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(6949..6979) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6994..7015 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7303..7891) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8062..8922) /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(8923..9027) /gene="bla" /label="AmpR promoter" primer_bind 9501..9517 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatga gcaggcacct ctggaagaat 2341 cagttgagtg agatggtgca acccagtggt ggcccagcag aggaccagga catgctgggt 2401 gaagaatctt ctctggggaa gccagcaatg ctacatcttc cttcagagca gggtactcct 2461 gagaccctcc agcgctgtct ggaagagaat caagaactcc gagatgctat ccggcagagc 2521 aatcagatgc tgagggaacg ctgtgaggag ctgctgcatt tccaggtcag ccagcgggag 2581 gagaaggagt tcctcatgtg caaattccag gaagccagga agctggtgga gagactgagc 2641 ttggagaagc ttgacctgcg gaggcagaga gaacaggcct tagaggattt ggagcatctg 2701 aagaaatgcc aacagcagat ggctgaggac aaggcctctg tgaaagctca ggtgacatca 2761 ttgctggggg aactccagga gagccagagt cgtttggagg ctgctaccaa ggaacgacaa 2821 actttagagg gaaggattcg agcagttagt gagcaggtca gacagctgga gagtgagcgg 2881 gaggtgctgc agcagcagca cagtgtccag gtggaccaac tgcgcatgca gaaccagagt 2941 gtggaggctg ccttgcgaat ggagcgacag gctgcttcag aggagaagcg gaagctggct 3001 cagttgcagg cagcctatca ccaactcttc caagactatg acagccacat taagagcagc 3061 aagggcatgc agctggaaga tctgaggcag cagctccagc aagctgagga ggccctggta 3121 gccaaacagg aattgattga taagctgaaa gaggaagctg agcagcacaa gattgtcatg 3181 gagaccgttc cagtcttgaa agcccaggcg gacatctaca aggctgactt ccaagctgag 3241 aggcatgccc gggagaaact ggtggagagg aaggagcttt tgcaggagca gttggagcag 3301 ctgcagcgcg agttcaacaa gttgaaagtt ggctgccatg agtcagccag gattgaggat 3361 atgaggaagc ggcatgtaga gacttcccag ccccctttac tccctgctcc agctcaccac 3421 tcctttcatt tggccttgtc caaccagcgg aggagccctc ctgaggaacc acctgacttc 3481 tgttgtccca agtgccagta tcaggctcct gatatggaca ctctacagat acatgtcatg 3541 gagtgcatag agggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3601 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3661 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3721 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3781 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3841 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3901 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3961 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4021 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4081 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4141 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4201 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4261 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4321 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4381 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4441 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4501 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4561 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4621 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4681 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4741 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4801 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4861 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4921 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 4981 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 5041 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5101 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5161 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5221 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5281 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5341 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5401 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5461 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5521 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5581 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5641 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5701 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5761 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5821 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5881 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5941 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 6001 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6061 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6121 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6181 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6241 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6301 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6361 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6421 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6481 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6541 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6601 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6661 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6781 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6841 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6901 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6961 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7021 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7081 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7141 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7201 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7261 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7321 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7381 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7441 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7501 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7561 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7621 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7681 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7741 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7801 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7861 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7921 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7981 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8041 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8101 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8161 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8221 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8281 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8341 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8401 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8461 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8521 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8581 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8641 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8701 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8761 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8821 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8881 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8941 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9001 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9061 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9121 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9181 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9241 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9301 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9361 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9421 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9481 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //