LOCUS V039857 9968 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039857 VERSION V039857 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..3805 /label="SLC7A11(NM_014331)" /note="SLC7A11(NM_014331)" /gene="SLC7A11" CDS 3812..3877 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3921..4132 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4145..4413 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4442..4451 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4472..5134 /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 5204..5257 /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 5264..6058 /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 6065..6653 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6725..6958 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7030..7151 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7170..7305 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7343..7359) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7367..7383 /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(7391..7421) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7436..7457 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7745..8333) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8504..9364) /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(9365..9469) /gene="bla" /label="AmpR promoter" primer_bind 9943..9959 /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 ccatggtcag aaagcctgtt gtgtccacca tctccaaagg 2341 aggttacctg cagggaaatg ttaacgggag gctgccttcc ctgggcaaca aggagccacc 2401 tgggcaggag aaagtgcagc tgaagaggaa agtcacttta ctgaggggag tctccattat 2461 cattggcacc atcattggag caggaatctt catctctcct aagggcgtgc tccagaacac 2521 gggcagcgtg ggcatgtctc tgaccatctg gacggtgtgt ggggtcctgt cactatttgg 2581 agctttgtct tatgctgaat tgggaacaac tataaagaaa tctggaggtc attacacata 2641 tattttggaa gtctttggtc cattaccagc ttttgtacga gtctgggtgg aactcctcat 2701 aatacgccct gcagctactg ctgtgatatc cctggcattt ggacgctaca ttctggaacc 2761 attttttatt caatgtgaaa tccctgaact tgcgatcaag ctcattacag ctgtgggcat 2821 aactgtagtg atggtcctaa atagcatgag tgtcagctgg agcgcccgga tccagatttt 2881 cttaaccttt tgcaagctca cagcaattct gataattata gtccctggag ttatgcagct 2941 aattaaaggt caaacgcaga actttaaaga cgccttttca ggaagagatt caagtattac 3001 gcggttgcca ctggcttttt attatggaat gtatgcatat gctggctggt tttacctcaa 3061 ctttgttact gaagaagtag aaaaccctga aaaaaccatt ccccttgcaa tatgtatatc 3121 catggccatt gtcaccattg gctatgtgct gacaaatgtg gcctacttta cgaccattaa 3181 tgctgaggag ctgctgcttt caaatgcagt ggcagtgacc ttttctgagc ggctactggg 3241 aaatttctca ttagcagttc cgatctttgt tgccctctcc tgctttggct ccatgaacgg 3301 tggtgtgttt gctgtctcca ggttattcta tgttgcgtct cgagagggtc accttccaga 3361 aatcctctcc atgattcatg tccgcaagca cactcctcta ccagctgtta ttgttttgca 3421 ccctttgaca atgataatgc tcttctctgg agacctcgac agtcttttga atttcctcag 3481 ttttgccagg tggcttttta ttgggctggc agttgctggg ctgatttatc ttcgatacaa 3541 atgcccagat atgcatcgtc ctttcaaggt gccactgttc atcccagctt tgttttcctt 3601 cacatgcctc ttcatggttg ccctttccct ctattcggac ccatttagta cagggattgg 3661 cttcgtcatc actctgactg gagtccctgc gtattatctc tttattatat gggacaagaa 3721 acccaggtgg tttagaataa tgtcggagaa aataaccaga acattacaaa taatactgga 3781 agttgtacca gaagaagata agttaggatc cgactacaaa gaccatgacg gtgattataa 3841 agatcatgac atcgactaca aggatgacga tgacaagtag cggccgcgaa ggatctgcga 3901 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 3961 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag 4021 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 4081 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct 4141 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg 4201 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt 4261 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct 4321 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt 4381 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga 4441 cgccaccatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac 4501 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccaagca 4561 gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct 4621 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga 4681 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta 4741 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat 4801 cggcgacttc aaggtggtgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa 4861 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgtgctggt 4921 gggcagcttc gcccgcacct tcagcctgcg cgacggcggc tactacagct tcgtggtgga 4981 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat 5041 gttcgccttc cgccgcgtgg aggagctgca cagcaacacc gagctgggca tcgtggagta 5101 ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc 5161 tgccgtggac ggcaccgccg gacccggctc caccggatct cgcgagggca gaggaagtct 5221 tctaacatgc ggtgacgtgg aggagaatcc cggccctgcc accatggctg aacaagatgg 5281 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 5341 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 5401 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 5461 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 5521 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 5581 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 5641 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 5701 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 5761 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 5821 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 5881 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 5941 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 6001 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagt 6061 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 6121 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 6181 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 6241 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 6301 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 6361 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 6421 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 6481 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 6541 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 6601 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcggtacct 6661 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 6721 ggactggaag ggctaattca ctcccaacga agataagatc tgctttttgc ttgtactggg 6781 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 6841 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 6901 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 6961 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 7021 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 7081 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 7141 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc 7201 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 7261 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag 7321 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 7381 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 7441 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 7501 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 7561 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 7621 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 7681 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 7741 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 7801 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 7861 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 7921 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7981 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 8041 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 8101 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 8161 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 8221 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 8281 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 8341 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 8401 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 8461 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 8521 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 8581 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 8641 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 8701 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 8761 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8821 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8881 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8941 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 9001 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 9061 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 9121 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 9181 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 9241 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 9301 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 9361 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 9421 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 9481 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 9541 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 9601 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 9661 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 9721 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 9781 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 9841 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 9901 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 9961 ccaagctg //