LOCUS V039394 9146 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039394 VERSION V039394 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 2303..2983 /label="YEATS4(NM_006530)" /note="YEATS4(NM_006530)" /gene="YEATS4" CDS 2990..3055 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3099..3310 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3323..3591 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3620..3629 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3650..4312 /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 4382..4435 /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 4442..5236 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5243..5831 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5903..6136 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6208..6329 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6348..6483 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6521..6537) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6545..6561 /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(6569..6599) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6614..6635 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6923..7511) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7682..8542) /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(8543..8647) /gene="bla" /label="AmpR promoter" primer_bind 9121..9137 /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 gaattcgcca ccatgttcaa gagaatggcc gaatttgggc ctgactccgg 2341 cgggagagta aagggtgtta ctatcgttaa accaatagtt tacggtaatg ttgctcggta 2401 ttttggaaag aaaagagaag aagatgggca cactcatcag tggacagtat atgtgaaacc 2461 atatagaaat gaggatatgt cagcatatgt gaagaaaatc cagtttaaat tacatgaaag 2521 ctatggcaat cctttaagag ttgttactaa acctccatat gaaattactg aaacaggatg 2581 gggtgaattc gaaataatca tcaaaatatt tttcattgac cctaatgaaa gacctgtaac 2641 cctgtatcat ttgctaaagc tgtttcaatc agacaccaat gcaatgctgg ggaaaaagac 2701 agtggtttca gagttctatg atgaaatgat atttcaagac ccaacagcaa tgatgcaaca 2761 attattgaca acatctcgtc agctaacatt aggagcctat aagcatgaaa cagaatttgc 2821 agagcttgaa gtgaaaacca gagaaaaatt agaagctgct aagaaaaaaa caagctttga 2881 gattgcagag cttaaggaga gattaaaagc aagtcgtgaa actataaatt gtttaaaaaa 2941 tgaaatcaga aaacttgaag aagatgacca agcaaaagac ataggatccg actacaaaga 3001 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg 3061 gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac 3121 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg 3181 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg 3241 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc 3301 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac 3361 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc 3421 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc 3481 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct 3541 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg 3601 accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga 3661 gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg 3721 agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct 3781 gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac 3841 ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa 3901 cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta 3961 cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag 4021 cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat 4081 gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta 4141 ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct 4201 gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga 4261 gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg 4321 cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg 4381 cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctgccac 4441 catggctgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 4501 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 4561 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 4621 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 4681 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 4741 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 4801 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 4861 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 4921 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga 4981 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 5041 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 5101 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 5161 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 5221 tgacgagttc ttctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac 5281 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 5341 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 5401 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 5461 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 5521 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 5581 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 5641 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 5701 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 5761 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 5821 cgcctccccg cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 5881 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag ataagatctg 5941 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc 6001 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg 6061 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg 6121 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca 6181 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa 6241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 6301 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa 6361 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 6421 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 6481 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt 6541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 6601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 6661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 6721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 6781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 6841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 6901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 6961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 7021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 7081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 7141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 7201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 7261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 7321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 7381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 7441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 7501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 7561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 7621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 7681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 7741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 7801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 7861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 7921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 7981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 8041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 8101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 8161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 8221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 8281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 8341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 8401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 8461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 8521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 8581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 8641 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 8701 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg 8761 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 8821 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct 8881 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 8941 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc 9001 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag 9061 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt 9121 gtaaaacgac ggccagtgcc aagctg //