LOCUS V032249 9550 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032249 VERSION V032249 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..3576 /label="NPFFR2(NM_004885)" /note="NPFFR2(NM_004885)" /gene="NPFFR2" CDS 3583..3648 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3692..3903 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3916..4184 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4213..4222 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4243..4905 /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 4975..5028 /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 5029..5628 /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 5629..6217 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6291..6524 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6596..6717 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6757..6892 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6925..6941) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6949..6965 /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(6973..7003) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7018..7039 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7327..7915) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8086..8946) /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(8947..9051) /gene="bla" /label="AmpR promoter" primer_bind 9525..9541 /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 atgagaaatg ggacacaaac 2341 tcttcagaaa actggcatcc catctggaat gtcaatgaca caaagcatca tctgtactca 2401 gatattaata ttacctatgt gaactactat cttcaccagc ctcaagtggc agcaatcttc 2461 attatttcct actttctgat cttctttttg tgcatgatgg gaaatactgt ggtttgcttt 2521 attgtaatga ggaacaaaca tatgcacaca gtcactaatc tcttcatctt aaacctggcc 2581 ataagtgatt tactagttgg catattctgc atgcctataa cactgctgga caatattata 2641 gcaggatggc catttggaaa cacgatgtgc aagatcagtg gattggtcca gggaatatct 2701 gtcgcagctt cagtctttac gttagttgca attgctgtag ataggttcca gtgtgtggtc 2761 taccctttta aaccaaagct cactatcaag acagcgtttg tcattattat gatcatctgg 2821 gtcctagcca tcaccattat gtctccatct gcagtaatgt tacatgtgca agaagaaaaa 2881 tattaccgag tgagactcaa ctcccagaat aaaaccagtc cagtctactg gtgccgggaa 2941 gactggccaa atcaggaaat gaggaagatc tacaccactg tgctgtttgc caacatctac 3001 ctggctcccc tctccctcat tgtcatcatg tatggaagga ttggaatttc actcttcagg 3061 gctgcagttc ctcacacagg caggaagaac caggagcagt ggcacgtggt gtccaggaag 3121 aagcagaaga tcattaagat gctcctgatt gtggccctgc tttttattct ctcatggctg 3181 cccctgtgga ctctaatgat gctctcagac tacgctgacc tttctccaaa tgaactgcag 3241 atcatcaaca tctacatcta cccttttgca cactggctgg cattcggcaa cagcagtgtc 3301 aatcccatca tttatggttt cttcaacgag aatttccgcc gtggtttcca agaagctttc 3361 cagctccagc tctgccaaaa aagagcaaag cctatggaag cttatgccct aaaagctaaa 3421 agccatgtgc tcataaacac atctaatcag cttgtccagg aatctacatt tcaaaaccct 3481 catggggaaa ccttgcttta taggaaaagt gctgaaaaac cccaacagga attagtgatg 3541 gaagaattaa aagaaactac taacagcagt gagattggat ccgactacaa agaccatgac 3601 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3661 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3721 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3781 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3841 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3901 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3961 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 4021 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 4081 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 4141 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 4201 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4261 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4321 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4381 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4441 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4501 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4561 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4621 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4681 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4741 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4801 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4861 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4921 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4981 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 5041 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5101 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5161 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5221 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5281 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5341 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5401 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5461 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5521 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5581 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5641 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5701 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5761 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5821 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5881 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5941 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6001 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 6061 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6121 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 6181 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6241 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6301 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6361 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6421 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6481 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6541 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6601 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6661 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6721 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6781 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6841 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6901 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7081 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7741 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8221 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9061 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9121 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9181 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9241 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9301 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9361 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9421 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9481 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9541 tgccaagctg //