LOCUS V042796 9511 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042796 VERSION V042796 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" regulatory 2279..2288 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2288..2359 /codon_start=1 /product="three tandem FLAG® epitope tags" /transl_table=1 /translation="DYKDDDDKDYKDDDDKDYKDDDDK" /label="3xFLAG" CDS 2366..3619 /label="Arrb1(NM_177231)" /note="Arrb1(NM_177231)" /gene="Arrb1" promoter 3670..3881 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3894..4162 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4191..4200 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4221..4883 /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 4953..5006 /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 5007..5606 /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 5607..6194 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6268..6501 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6573..6694 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6713..6848 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6886..6902) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6910..6926 /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(6934..6964) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6979..7000 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7288..7876) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8047..8907) /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(8908..9012) /gene="bla" /label="AmpR promoter" primer_bind 9486..9502 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 caccatggac tacaaggatg acgatgacaa ggattacaaa gacgacgatg ataaggacta 2341 taaggatgat gacgacaaag aattcatggg cgacaaaggg acacgagtgt tcaagaaggc 2401 aagccccaat ggaaagctca ctgtctacct gggaaagcgg gactttgtgg accacattga 2461 cctcgtggac cccgtggatg gtgtggtcct ggtggatcct gagtatctca aagaaaggcg 2521 agtctacgtg acactgacct gcgccttccg gtatggccgg gaagacctgg atgtcttggg 2581 tctgactttt cggaaagacc tgtttgtggc caatgtgcag tccttcccac cggcccccga 2641 ggacaagaag ccactgacac ggctacagga gcgactcatc aagaagctgg gcgagcatgc 2701 ctgccccttc acctttgaga tcccgccaaa ccttccatgc tcagtcactc tgcagcctgg 2761 gcctgaggac acaggaaagg cctgtggtgt ggattatgaa gtcaaagcct tctgtgctga 2821 gaacctggag gagaagatcc acaaaaggaa ctctgtgcgg ctagtcatcc ggaaggttca 2881 gtatgcccct gagagacctg gccctcagcc cacggctgag actaccagac agttccttat 2941 gtcagacaag cccctacacc ttgaggcatc actggataag gagatctatt accacggaga 3001 acccatcagc gttaatgtcc acgtcaccaa caacaccaac aagactgtga agaagatcaa 3061 gatctcggtg cgccagtatg cagacatctg tctcttcaac acagctcagt acaagtgccc 3121 agtggccatg gaggaggctg atgacaatgt ggcacccagc tcaacattct gcaaggtcta 3181 tacgctgact cccttcctgg ccaacaaccg agagaagcgg ggactcgccc tcgacgggaa 3241 gctcaagcat gaggacacga atctggcttc cagcacgctg ttgcgggaag gtgccaaccg 3301 tgaaatcctg ggcatcatcg tttcctacaa agtcaaggtg aaactggtgg tgtcccgggg 3361 cggcctcttg ggagaccttg catccagtga tgtggccgtg gagctgcctt ttaccctaat 3421 gcaccccaag cctaaggagg agcccccaca tcgggaagtt ccagagagcg agactccagt 3481 agacaccaat ctcatagagc ttgacaccaa tgatgacgac attgtatttg aggactttgc 3541 tcgccagcgg ctgaaaggca tgaaggatga caaggacgaa gaggatgacg gcaccggctc 3601 tccgcacctc aacaacagat agggatccgc ggccgcgaag gatctgcgat cgctccggtg 3661 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 3721 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 3781 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 3841 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc 3901 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc 3961 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt 4021 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca 4081 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt 4141 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac gccaccatgg 4201 agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga 4261 acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag ggccgcatga 4321 ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg 4381 tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc 4441 tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg 4501 gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca 4561 aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca 4621 gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg 4681 cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac agccacatgc 4741 acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc 4801 gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac cagcacgcct 4861 tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct gccgtggacg 4921 gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt ctaacatgcg 4981 gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca 5041 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 5101 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 5161 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 5221 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 5281 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 5341 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 5401 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 5461 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 5521 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 5581 gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat 5641 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 5701 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 5761 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 5821 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 5881 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 5941 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 6001 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt 6061 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 6121 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctcccttt 6181 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta 6241 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag 6301 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 6361 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 6421 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6481 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 6541 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt 6601 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 6661 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc 6721 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6781 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6841 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc 6901 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6961 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 7021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 7081 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 8041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 8101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 8161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 8221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8341 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8401 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8461 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8521 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8581 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8641 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8701 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8761 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8821 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8881 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8941 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 9001 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 9061 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 9121 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 9181 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 9241 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 9301 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 9361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 9421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 9481 acgttgtaaa acgacggcca gtgccaagct g //