LOCUS V026616 9610 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026616 VERSION V026616 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..3636 /label="GPR180(NM_180989)" /note="GPR180(NM_180989)" /gene="GPR180" CDS 3643..3708 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3752..3963 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3976..4244 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4273..4282 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4303..4965 /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 5035..5088 /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 5089..5688 /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 5689..6277 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6351..6584 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6656..6777 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6817..6952 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6985..7001) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7009..7025 /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(7033..7063) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7078..7099 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7387..7975) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8146..9006) /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(9007..9111) /gene="bla" /label="AmpR promoter" primer_bind 9585..9601 /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 gccaccatgg gggggctgcg gctgctggct 2341 gtggccctca cgtgctgctg gtggccgcag ggcagccagg gtaagaccct gcggggcagc 2401 ttcagcagca ccgcggccca ggacgcccag ggccagcgca tcggccactt cgagttccat 2461 ggtgaccatg ctcttctgtg tgtcagaatc aacaacatag cagtagctgt tggaaaagaa 2521 gctaaactct acctgttcca agcccaggaa tggctaaagc tacagcaaag cagtcatggt 2581 tatagctgta gtgaaaaatt atccaaagct cagttgacaa tgaccatgaa ccagaccgaa 2641 cataatctga cagtgtccca gattccgtct ccacaaacgt ggcatgtgtt ttatgcagac 2701 aagtatacat gccaagatga caaggagaat tctcaggtgg aagatatccc atttgaaatg 2761 gtgttactaa acccagatgc cgaagggaat ccatttgatc attttagtgc tggagaatct 2821 gggttacatg agttcttttt cctcctagtc ctagtgtact ttgtgattgc ttgcatttat 2881 gctcaatcat tgtggcaggc tattaagaaa ggcggaccca tgcacatgat tttaaaggtt 2941 ctgacaactg cattgctgtt acaagctggt tcagctttag ctaattacat tcatttctcc 3001 agttactcca aagatggaat aggggtacca tttatgggaa gtttggcaga attttttgac 3061 atcgcttccc aaattcagat gttatactta cttttgagtc tatgcatggg ttggacaata 3121 gtcagaatga agaagtctca aagcagacct ctccagtggg attctacgcc tgcatccact 3181 ggcattgcag tattcattgt catgacacag agtgttttgc tactttggga acagtttgaa 3241 gatatcagtc atcatagcta ccattcacac cacaacttag cagggatcct cctaattgtt 3301 ctaagaattt gcctagcatt gtcattaggc tgtggactct atcagatcat cacagtggag 3361 agaagtacac tcaaaaggga gttctacatc acatttgcca aaggctgtat cttgtggttt 3421 ttatgccatc cagttcttgc atgcatttct gtcattttta gcgactacca aagagacaag 3481 gttattacaa taggtgttat cctttgccag tctgtttcca tggttattct ctacagactc 3541 tttctgtctc acagtctata ctgggaagtt tcttcacttt cttcagtaac actaccactg 3601 accatatcat ctggacacaa aagtcgccct catttcggat ccgactacaa agaccatgac 3661 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3721 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3781 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3841 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3901 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3961 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 4021 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 4081 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 4141 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 4201 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 4261 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 4321 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 4381 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 4441 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 4501 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 4561 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4621 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4681 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4741 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4801 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4861 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4921 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4981 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 5041 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 5101 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5161 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5221 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5281 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5341 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5401 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5461 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5521 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5581 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5641 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5701 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5761 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5821 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5881 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5941 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 6001 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 6061 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 6121 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 6181 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 6241 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 6301 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 6361 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 6421 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6481 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6541 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6601 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6661 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6721 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6781 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6841 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6901 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6961 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 7021 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 7081 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7141 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7201 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7261 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7321 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7801 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7921 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 8041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 9001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9121 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 9181 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 9241 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 9301 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9361 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9421 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9481 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9541 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9601 tgccaagctg //