LOCUS V034754 9686 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034754 VERSION V034754 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2290..2985 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /transl_table=1 /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" /label="mKate2" CDS 3022..4161 /label="Dek(NM_025900)" /note="Dek(NM_025900)" /gene="Dek" CDS 4162..4179 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 4229..4440 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4453..4721 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4749..5774 /codon_start=1 /gene="aph(4)-Ia" /note="confers resistance to hygromycin" /product="aminoglycoside phosphotransferase from E. coli" /transl_table=1 /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E*" /label="HygR" misc_feature 5781..6369 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6443..6676 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6748..6869 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6888..7023 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7061..7077) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7085..7101 /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(7109..7139) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7154..7175 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7463..8051) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8222..9082) /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(9083..9187) /gene="bla" /label="AmpR promoter" primer_bind 9661..9677 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagccacca tggtgagcga gctgattaag gagaacatgc acatgaagct gtacatggag 2341 ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag 2401 ggcacccaga ccatgagaat caaggcggtc gagggcggcc ctctcccctt cgccttcgac 2461 atcctggcta ccagcttcat gtacggcagc aaaaccttca tcaaccacac ccagggcatc 2521 cccgacttct ttaagcagtc cttccccgag ggcttcacat gggagagagt caccacatac 2581 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc 2641 tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa 2701 acactcggct gggaggcctc caccgagacc ctgtaccccg ctgacggcgg cctggaaggc 2761 agagccgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttgaagacc 2821 acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac 2881 agaagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg 2941 gctgtggcca gatactgcga cctccctagc aaactggggc acagaggtgg aggcggatca 3001 ggaggcggtg gatcttctag aatgtcggcg gcggcggccc ccgctgcgga gggagaggac 3061 gcccccgtgc cgccctcatc cgagaaggaa cccgagatgc cgggtcccag ggaagagagt 3121 gaggaggagg aggaggatga cgaagacgat gatgaagagg acgaggagga agaaaaagaa 3181 aagagtctta tcgtggaagg caagagagag aagaagaaag tagagagact gacgatgcaa 3241 gtgtcttcct tacagagaga gccatttaca gtgacacaag ggaagggtca gaaactttgt 3301 gaaattgaaa ggatacattt ctttctgagt aagaaaaaac cagatgaact tagaaatcta 3361 cacaaactgc tttacaacag gccgggcaca gtgtcctcgt tgaagaagaa cgtgggtcag 3421 ttcagtggct ttccattcga aaaaggcagt acccagtata aaaagaagga agaaatgttg 3481 aaaaagtttc gaaatgccat gttaaagagc atctgtgagg ttcttgattt agagaggtca 3541 ggcgtgaaca gcgaactcgt gaagaggatc ttgaacttct taatgcatcc aaagccttct 3601 ggcaaaccat taccaaaatc caaaaaatct tccagcaaag gtagtaaaaa ggaacggaac 3661 agttctggaa caacaaggaa gtcaaagcaa actaaatgcc ctgaaattct gtcagatgag 3721 tctagtagtg atgaagatga gaagaaaaat aaggaagagt cttcggaaga tgaagagaaa 3781 gaaagtgaag aggagcaacc accaaaaaag acatctaaaa aagaaaaagc aaaacagaaa 3841 gctactgcta aaagtaaaaa atctgtgaag agtgctaatg ttaagaaggc agacagcagt 3901 accaccaaga agaatcaaaa aagttccaaa aaagagtctg aatctgaaga cagttctgat 3961 gatgaaccct taattaaaaa attgaaaaag ccacctacag atgaagagct aaaggaaaca 4021 gtgaagaaat tactggctga tgctaacttg gaagaagtca caatgaagca gatttgcaaa 4081 gaggtatatg aaaattatcc tgcttatgat ttgactgaga ggaaagattt cattaaaaca 4141 actgtaaaag agctaatttc tcatcaccat catcaccatt gaggatccgc ggccgcaagg 4201 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag 4261 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac 4321 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat 4381 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag 4441 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc 4501 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg 4561 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct 4621 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc 4681 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct 4741 acgtcgacat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa 4801 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca 4861 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct 4921 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc 4981 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 5041 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg 5101 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac 5161 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc 5221 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc 5281 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg 5341 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga 5401 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt 5461 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag 5521 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct 5581 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc 5641 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga 5701 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga 5761 gggcaaagga ataggtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg 5821 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 5881 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 5941 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 6001 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 6061 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 6121 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat 6181 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 6241 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 6301 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 6361 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6421 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg 6481 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc 6541 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg 6601 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg 6661 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca 6721 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa 6781 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 6841 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa 6901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 6961 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 7021 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt 7081 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 7141 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 7201 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 7261 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 7321 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 7381 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 7441 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 7501 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 7561 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 7621 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 7681 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 7741 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 7801 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 7861 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 7921 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 7981 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 8041 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 8101 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 8161 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 8221 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 8281 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 8341 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 8401 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 8461 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 8521 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 8581 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 8641 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 8701 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 8761 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 8821 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 8881 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 8941 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 9001 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 9061 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 9121 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 9181 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 9241 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg 9301 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 9361 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct 9421 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 9481 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc 9541 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag 9601 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt 9661 gtaaaacgac ggccagtgcc aagctg //