LOCUS V036711 9718 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036711 VERSION V036711 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..3744 /label="Nsun6(NM_028950)" /note="Nsun6(NM_028950)" /gene="Nsun6" CDS 3751..3816 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3860..4071 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4084..4352 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4381..4390 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4411..5073 /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 5143..5196 /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 5197..5796 /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 5797..6385 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6459..6692 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6764..6885 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6925..7060 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7093..7109) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7117..7133 /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(7141..7171) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7186..7207 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7495..8083) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8254..9114) /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(9115..9219) /gene="bla" /label="AmpR promoter" primer_bind 9693..9709 /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 gccaccatgt atgtatttcc aaagatatct 2341 ttgagacctg aagttgaaaa ctatctcaaa gagagctttt tgaacgagga ggctgtgtct 2401 gcttcaagca gacaagaagc agaaaggaaa tttgagactc tgctgcttcg cctgtcacac 2461 cctccgtcga tgacaactgt cagagtgaat acgcatttag gctcagttca gcatgtcaga 2521 ggtctgctgc ttgaagagct ccagaagcag tttggtgaat ccagcattcc tgttgttcag 2581 catccagccc tcccggatgt cttgctcatc cccatgactg gaccaaggaa gaatatagaa 2641 agacaacagg gtgaagtgat tgttggcgca cagtgtggca atgcagtgtt aagaggggcc 2701 catgtctatg ttccaggaat tgtgtcagct tcaaaattta tgaaagctgg agatgttatt 2761 tccgtgtatt ctgatattaa cggcaaatgt aagaaaggag ccaaagaatt tgatggaact 2821 aaagtatttc tcggaaacgg aatttctgaa ctgagccgca aagatatctt caatgggtta 2881 cctgatctga aaggtatagg cataagaatg acagaaccca tatatctcag tccctcgttt 2941 gacaatgttc tacccagtta tatattttta cagaatttac catctacagt tgtagctcat 3001 gttcttgatc ctcaacctgg agagaagatt ctagatatgt gtgcagcccc tgggggcaaa 3061 actacacata ctgcagcact gatgcaggac aagggagaag taatagcatt ggacaaaata 3121 ttgacaaaag tgaacaaact gaagcagaat gcttcattat tagggcttca ttctatccgg 3181 gcattttgct tcgatgcaac aaaggcactt aagcttgata ctaccgatgg catagaaggg 3241 gggcctccat tcttaccaga atcttttgac cgaattatcc ttgatgcacc ctgcagtgga 3301 atgggacaga ggccaaatat ggcttgtact tggactttaa aggaagtgac atcctatcag 3361 ccactacagc gaaaactcct tcatgtggca gttcaactcc tgaagccagg gggagtgctg 3421 gtctacagca catgcacaat aaccttggca gaaaatgaag agcaagttgc ctgggccctc 3481 cgaacattcc cctgccttca gcttcaaccc caggaaccac agattggagg agaagggatg 3541 gtgggagctg gcctgacatt ggagcaattg aagcagctgc agcgatttga tccgtctgtt 3601 gtaccactgc agaacatgga cactgattct cttggagagg ccagaaggga agacatgatt 3661 tggttggcca acaaagactg tataggcttc tttattgcaa agtttctaaa atgtcaaagc 3721 acaaaagcga aggtttctca gaaaggatcc gactacaaag accatgacgg tgattataaa 3781 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat 3841 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3901 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3961 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 4021 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4081 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4141 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4201 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4261 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4321 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 4381 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc 4441 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag 4501 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg 4561 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag 4621 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac 4681 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc 4741 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag 4801 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg 4861 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac 4921 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg 4981 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac 5041 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct 5101 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt 5161 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg 5221 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 5281 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 5341 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 5401 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 5461 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 5521 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 5581 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 5641 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 5701 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg 5761 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt 5821 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5881 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5941 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 6001 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 6061 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 6121 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 6181 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg 6241 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6301 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6361 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc 6421 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 6481 acgaagataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 6541 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 6601 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 6661 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca 6721 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 6781 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6841 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 6901 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6961 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 7021 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa 7081 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 7141 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 7201 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 7261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 7321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 7381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 7441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 7501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 7561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 7621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 7681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 7741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 7801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 7861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 7921 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 7981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 8041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 8101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 8161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 8221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 8281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 8341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 8401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 8461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 8521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 8581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 8641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 8701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 8761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 8821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 8881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 8941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 9001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 9061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 9121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 9181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9241 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 9301 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 9361 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 9421 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 9481 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 9541 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 9601 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 9661 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //