LOCUS V038812 9667 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038812 VERSION V038812 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..3693 /label="Htr2c(NM_008312)" /note="Htr2c(NM_008312)" /gene="Htr2c" CDS 3700..3765 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3809..4020 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4033..4301 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4330..4339 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4360..5022 /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 5092..5145 /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 5146..5745 /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 5746..6334 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6408..6641 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6713..6834 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6874..7009 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7042..7058) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7066..7082 /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(7090..7120) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7135..7156 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7444..8032) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8203..9063) /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(9064..9168) /gene="bla" /label="AmpR promoter" primer_bind 9642..9658 /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 tgaacctggg cactgcggtg 2341 cgctcactcc ttgtgcacct aattggccta ttggtttggc agttcgatat ttccataagt 2401 ccagtagcag ctatagtaac tgacactttt aattcctccg atggtggacg cttgtttcaa 2461 ttcccggacg gggtacaaaa ctggccagca ctttcaatag tcgtgattat aatcatgaca 2521 atagggggca acattctcgt tatcatggca gtaagcatgg agaagaaact gcacaatgct 2581 accaattatt tcttaatgtc cctagccatt gctgatatgc tggtgggact acttgtcatg 2641 cccctgtctc tgcttgcaat tctttatgat tatgtctggc ctttacctag atatttgtgc 2701 cccgtctgga tttcactaga tgtgctattt tcaactgcgt ccatcatgca cctctgcgcc 2761 atatcgctgg accggtatgt agcaatacgt aatcctattg agcatagccg gttcaattcg 2821 cggactaagg ccatcatgaa gattgccatc gtttgggcaa tatcaatagg agtttcagtt 2881 cctatccctg tgattggact gagggacgaa agcaaagtgt tcgtgaataa tactacctgc 2941 gtgctcaatg acccgaactt cgttctcatc gggtccttcg tggcattctt catcccgttg 3001 acaattatgg tgatcaccta cttcttaacg atctacgtcc tacgccgtca aaccctgatg 3061 ttacttcgag gtcacaccga ggaggaactg cgtaatatca gcctgaactt tctaaagtgc 3121 tgctgcaaga agggtgatga ggaagagaac gctcccaacc ccaatccaga tcagaagcca 3181 cgtcgaaaga agaaagaaaa gcggcctaga ggcaccatgc aagctatcaa caatgagaag 3241 aaagcttcca aagtccttgg cattgtattc tttgtgtttc tgatcatgtg gtgcccgttt 3301 ttcatcacca atatcctgtc ggtgctttgt gggaaggcct gtaaccaaaa gctaatggag 3361 aaacttctca atgtgtttgt ttggattggc tatgtgtgtt caggcatcaa tcctctggtg 3421 tacactctct tcaacaaaat ttaccgaagg gctttctcta aatatttgcg ctgcgattat 3481 aagccagaca aaaagcctcc tgttcgacag attcctaggg ttgctgccac tgctttgtct 3541 gggagggagc tcaatgttaa catttatcgg cataccaatg aacgtgtagt taggaaagct 3601 aatgacacag agcctggcat agagatgcag gtagagaatt tagagctgcc ggtcaatccc 3661 tctaatgtgg tcagcgagag gattagtagt gtgggatccg actacaaaga ccatgacggt 3721 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg 3781 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 3841 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 3901 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 3961 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4021 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4081 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4141 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4201 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4261 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4321 acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc 4381 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc 4441 cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc 4501 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc 4561 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc 4621 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc 4681 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc 4741 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac 4801 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc 4861 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg 4921 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc 4981 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg 5041 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga 5101 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag 5161 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 5221 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 5281 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 5341 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 5401 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 5461 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 5521 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 5581 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 5641 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 5701 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat 5761 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5821 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5881 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5941 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 6001 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6061 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6121 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 6181 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6241 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6301 acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta 6361 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6421 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca 6481 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 6541 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 6601 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct 6661 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt 6721 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6781 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6841 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 6901 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 6961 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc 7021 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 7081 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 7141 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 7201 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 7261 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7321 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7381 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7441 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7501 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7561 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7621 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7681 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7741 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7801 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7861 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7921 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7981 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 8041 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 8101 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 8161 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8221 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8281 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8341 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8401 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8461 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8521 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8581 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8641 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8701 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8761 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8821 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8881 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8941 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 9001 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 9061 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 9121 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 9181 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 9241 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 9301 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 9361 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 9421 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 9481 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9541 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9601 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9661 caagctg //