LOCUS V019296 9447 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019296 VERSION V019296 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2297..3532 /label="ASS1(NM_000050)" /note="ASS1(NM_000050)" /gene="ASS1" CDS 3533..3556 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3606..3817 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3830..4098 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4127..4136 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4157..4819 /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 4889..4942 /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 4943..5542 /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 5543..6130 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6204..6437 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6509..6630 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6649..6784 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6822..6838) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6846..6862 /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(6870..6900) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6915..6936 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7224..7812) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7983..8843) /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(8844..8948) /gene="bla" /label="AmpR promoter" primer_bind 9422..9438 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgaattc gccaccatgt ccagcaaagg ctccgtggtt ctggcctaca gtggcggcct 2341 ggacacctcg tgcatcctcg tgtggctgaa ggaacaaggc tatgacgtca ttgcctatct 2401 ggccaacatt ggccagaagg aagacttcga ggaagccagg aagaaggcac tgaagcttgg 2461 ggccaaaaag gtgttcattg aggatgtcag cagggagttt gtggaggagt tcatctggcc 2521 ggccatccag tccagcgcac tgtatgagga ccgctacctc ctgggcacct ctcttgccag 2581 gccctgcatc gcccgcaaac aagtggaaat cgcccagcgg gagggggcca agtatgtgtc 2641 ccacggcgcc acaggaaagg ggaacgatca ggtccggttt gagctcagct gctactcact 2701 ggccccccag ataaaggtca ttgctccctg gaggatgcct gaattctaca accggttcaa 2761 gggccgcaat gacctgatgg agtacgcaaa gcaacacggg attcccatcc cggtcactcc 2821 caagaacccg tggagcatgg atgagaacct catgcacatc agctacgagg ctggaatcct 2881 ggagaacccc aagaaccaag cgcctccagg tctctacacg aagacccagg acccagccaa 2941 agcccccaac acccctgaca ttctcgagat cgagttcaaa aaaggggtcc ctgtgaaggt 3001 gaccaacgtc aaggatggca ccacccacca gacctccttg gagctcttca tgtacctgaa 3061 cgaagtcgcg ggcaagcatg gcgtgggccg tattgacatc gtggagaacc gcttcattgg 3121 aatgaagtcc cgaggtatct acgagacccc agcaggcacc atcctttacc atgctcattt 3181 agacatcgag gccttcacca tggaccggga agtgcgcaaa atcaaacaag gcctgggctt 3241 gaaatttgct gagctggtgt ataccggttt ctggcacagc cctgagtgtg aatttgtccg 3301 ccactgcatc gccaagtccc aggagcgagt ggaagggaaa gtgcaggtgt ccgtcctcaa 3361 gggccaggtg tacatcctcg gccgggagtc cccactgtct ctctacaatg aggagctggt 3421 gagcatgaac gtgcagggtg attatgagcc aactgatgcc accgggttca tcaacatcaa 3481 ttccctcagg ctgaaggaat atcatcgtct ccagagcaag gtcactgcca aagattacaa 3541 ggatgacgac gataagtgag gatccgcggc cgcaaggatc tgcgatcgct ccggtgcccg 3601 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa 3661 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg 3721 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa 3781 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat 3841 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt 3901 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa 3961 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg 4021 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt 4081 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagacgcca ccatggagag 4141 cgacgagagc ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg 4201 cgtggagttc gagctggtgg gcggcggaga gggcaccccc aagcagggcc gcatgaccaa 4261 caagatgaag agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat 4321 gggctacggc ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca 4381 cgccatcaac aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt 4441 gctgcacgtg agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt 4501 ggtgggcacc ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa 4561 cgccaccgtg gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg 4621 caccttcagc ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt 4681 caagagcgcc atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg 4741 cgtggaggag ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa 4801 gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg tggacggcac 4861 cgccggaccc ggctccaccg gatctcgcga gggcagagga agtcttctaa catgcggtga 4921 cgtggaggag aatcccggcc ctatgaccga gtacaagccc acggtgcgcc tcgccacccg 4981 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 5041 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt 5101 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 5161 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 5221 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 5281 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga 5341 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 5401 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg 5461 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 5521 gacccgcaag cccggtgcct gaaatcaacc tctggattac aaaatttgtg aaagattgac 5581 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 5641 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 5701 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 5761 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 5821 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 5881 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct 5941 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 6001 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 6061 tctgcggcct cttccgcgtc tccgccttcg ccctcagacg agtcggatct ccctttggcc 6121 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 6181 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct 6241 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 6301 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 6361 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 6421 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 6481 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa 6541 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 6601 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta 6661 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 6721 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 6781 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg 6841 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6901 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6961 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 7021 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 7081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 7141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 7201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 7261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 7321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 7381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 7441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 7501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7621 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 7681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 8041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 8101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 8161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 8221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 8281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 8341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 8401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 8461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 8521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 8581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 8761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8941 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 9001 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 9061 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 9121 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 9181 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 9241 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg 9301 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 9361 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 9421 tgtaaaacga cggccagtgc caagctg //