LOCUS V044978 10031 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044978 VERSION V044978 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..4125 /label="Plk1(NM_017100)" /note="Plk1(NM_017100)" /gene="Plk1" promoter 4190..4401 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4414..4682 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4711..4720 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4741..5403 /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 5473..5526 /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 5527..6126 /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 6127..6714 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6788..7021 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7093..7214 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7233..7368 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7406..7422) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7430..7446 /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(7454..7484) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7499..7520 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7808..8396) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8567..9427) /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(9428..9532) /gene="bla" /label="AmpR promoter" primer_bind 10006..10022 /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 gccaccatga atgcagcggc caaagctgga 2341 aagctggctc gagcaccagc cgacctaggg aaaggtgggg tcccgggaga cgcagctccc 2401 ggtgccccag gggccgcccc gctggcgaaa gagattccgg aggtcctgat ggatccacgc 2461 agccggcggc agtatgtacg gggccgcttt ctgggtaaag gaggcttcgc caaatgcttc 2521 gaaatctcag actcagacac aaaagaggtg ttcgcaggca agatcgtgcc taagtctttg 2581 ctgctcaagc cccatcagaa ggagaaaatg tctatggaga tctccattca ccgcagcctc 2641 gcacaccagc acgtcgtagg cttccatggc ttttttgagg acagcgactt tgtgttcgtg 2701 gttttggagc tctgtcgcag gaggtctctc ctggagctgc acaaaaggag gaaggcgctg 2761 accgagcccg aggcccgata ctacctgcgg cagatagtcc tgggttgcca gtacctgcac 2821 cgcaatcaag tcattcacag ggacctcaaa ctgggcaacc tctttctgaa tgaggatctg 2881 gaggtgaaaa taggggattt tggcctggca acaaaagtgg aatatgaagg ggaacggaag 2941 aagaccctgt gtggcactcc taactacata gctcccgagg tgctgagcaa gaagggacac 3001 agctttgagg tggacgtgtg gtccattggg tgcatcatgt ataccttgct ggtgggcaag 3061 ccaccttttg agacctcatg cctaaaagag acctacctcc ggatcaagaa aaatgaatac 3121 agtattccca agcacatcaa cccagtggct gcctccctca tccagaaaat gcttcagaca 3181 gaccccactg cccgccccac cattcacgag ttgctcaatg acgagttctt cacttccggc 3241 tacatccccg cccgtctgcc tattacctgc ctcaccatcc caccaaggtt ttcaatagct 3301 cccagcagcc tagaccccag caacaggaag cctctcacag tccttaataa aggtgtggag 3361 aaccccctgc ctgaccgtcc ccgagagaaa gaggaaccag tggttcggga gacaaatgag 3421 gccatcgagt gccaccttag tgacttgcta cagcagctga ccagtgtcaa cgcctccaag 3481 ccctcagagc gagggctggt gcggcaagag gaggccgagg atcctgcttg catccctatc 3541 ttctgggtca gcaaatgggt agactattcg gacaagtatg gccttggtta tcagctatgt 3601 gacaacagtg tgggagtgct ctttaatgac tcaacacgcc tcatcctcta caatgacggg 3661 gacagcctgc agtacataga gcgtgatggc acggagtcct acctcactgt gagctcccac 3721 cccaactcct tgatgaagaa gatcactctc ctcaactatt tccgcaatta catgagtgaa 3781 cacctgttga aggcaggggc caacatcacg ccccgggaag gcgacgagct ggcccggctg 3841 ccctacctac gaacatggtt ccgcacacgc agcgccatca tcctgcacct cagcaatggc 3901 acagtacaga ttaatttctt ccaggaccac accaaactta tcctgtgccc cctgatggca 3961 gcagtgacct acatcaacga gaagagggac ttccggacat accgcctgag ccttctggag 4021 gaatatggct gctgcaagga actggctagc cgcctccgct atgcccgcac catggtagac 4081 aagctgctga gctcgcgttc cgcctgcaac cgcctcaagg cctcctaggg atccctgcga 4141 tatccagcgc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc 4201 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag 4261 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc 4321 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac 4381 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc 4441 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc 4501 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg 4561 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc 4621 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga 4681 tccaagctgt gaccggcgcc tacgctagac gccaccatgg agagcgacga gagcggcctg 4741 cccgccatgg agatcgagtg ccgcatcacc ggcaccctga acggcgtgga gttcgagctg 4801 gtgggcggcg gagagggcac ccccaagcag ggccgcatga ccaacaagat gaagagcacc 4861 aaaggcgccc tgaccttcag cccctacctg ctgagccacg tgatgggcta cggcttctac 4921 cacttcggca cctaccccag cggctacgag aaccccttcc tgcacgccat caacaacggc 4981 ggctacacca acacccgcat cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc 5041 agctaccgct acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg caccggcttc 5101 cccgaggaca gcgtgatctt caccgacaag atcatccgca gcaacgccac cgtggagcac 5161 ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc 5221 gacggcggct actacagctt cgtggtggac agccacatgc acttcaagag cgccatccac 5281 cccagcatcc tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga ggagctgcac 5341 agcaacaccg agctgggcat cgtggagtac cagcacgcct tcaagacccc catcgccttc 5401 gccagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg acccggctcc 5461 accggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc 5521 ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg 5581 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat 5641 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg 5701 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg 5761 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg 5821 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc 5881 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt 5941 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc 6001 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc 6061 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt 6121 gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 6181 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 6241 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 6301 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 6361 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 6421 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 6481 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 6541 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 6601 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 6661 cgtctccgcc ttcgccctca gacgagtcgg atctcccttt ggccgcctcc ccgcctggta 6721 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 6781 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact 6841 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 6901 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 6961 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 7021 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca 7081 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 7141 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 7201 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg 7261 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 7321 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca 7381 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 7441 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 7501 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 7561 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 7621 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 7681 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 7741 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 7801 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 7861 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 7921 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 7981 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 8041 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 8101 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 8161 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 8221 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 8281 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 8341 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 8401 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 8461 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 8521 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 8581 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 8641 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 8701 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 8761 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 8821 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 8881 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 8941 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 9001 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 9061 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 9121 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 9181 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 9241 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 9301 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 9361 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 9421 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 9481 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 9541 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 9601 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct 9661 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 9721 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg 9781 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg 9841 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag 9901 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 9961 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 10021 gtgccaagct g //