LOCUS V038272 9250 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038272 VERSION V038272 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 2302..3357 /label="WNT3A(NM_033131)" /note="WNT3A(NM_033131)" /gene="WNT3A" promoter 3408..3619 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3632..3900 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3929..3938 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3959..4621 /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 4691..4744 /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 4745..5344 /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 5345..5933 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6007..6240 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6312..6433 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6452..6587 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6625..6641) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6649..6665 /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(6673..6703) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6718..6739 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7027..7615) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7786..8646) /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(8647..8751) /gene="bla" /label="AmpR promoter" primer_bind 9225..9241 /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 agagctagcg aattcgccac catggcccca ctcggatact tcttactcct ctgcagcctg 2341 aagcaggctc tgggcagcta cccgatctgg tggtcgctgg ctgttgggcc acagtattcc 2401 tccctgggct cgcagcccat cctgtgtgcc agcatcccgg gcctggtccc caagcagctc 2461 cgcttctgca ggaactacgt ggagatcatg cccagcgtgg ccgagggcat caagattggc 2521 atccaggagt gccagcacca gttccgcggc cgccggtgga actgcaccac cgtccacgac 2581 agcctggcca tcttcgggcc cgtgctggac aaagctacca gggagtcggc ctttgtccac 2641 gccattgcct cagccggtgt ggcctttgca gtgacacgct catgtgcaga aggcacggcc 2701 gccatctgtg gctgcagcag ccgccaccag ggctcaccag gcaagggctg gaagtggggt 2761 ggctgtagcg aggacatcga gtttggtggg atggtgtctc gggagttcgc cgacgcccgg 2821 gagaaccggc cagatgcccg ctcagccatg aaccgccaca acaacgaggc tgggcgccag 2881 gccatcgcca gccacatgca cctcaagtgc aagtgccacg ggctgtcggg cagctgcgag 2941 gtgaagacat gctggtggtc gcaacccgac ttccgcgcca tcggtgactt cctcaaggac 3001 aagtacgaca gcgcctcgga gatggtggtg gagaagcacc gggagtcccg cggctgggtg 3061 gagaccctgc ggccgcgcta cacctacttc aaggtgccca cggagcgcga cctggtctac 3121 tacgaggcct cgcccaactt ctgcgagccc aaccctgaga cgggctcctt cggcacgcgc 3181 gaccgcacct gcaacgtcag ctcgcacggc atcgacggct gcgacctgct gtgctgcggc 3241 cgcggccaca acgcgcgagc ggagcggcgc cgggagaagt gccgctgcgt gttccactgg 3301 tgctgctacg tcagctgcca ggagtgcacg cgcgtctacg acgtgcacac ctgcaagtag 3361 ggatccgcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac 3421 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag 3481 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 3541 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 3601 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg 3661 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg 3721 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg 3781 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct 3841 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc 3901 caagctgtga ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc 3961 cgccatggag atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt 4021 gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa 4081 aggcgccctg accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca 4141 cttcggcacc taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg 4201 ctacaccaac acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag 4261 ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc 4321 cgaggacagc gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct 4381 gcaccccatg ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga 4441 cggcggctac tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc 4501 cagcatcctg cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag 4561 caacaccgag ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc 4621 cagatcccgc gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac 4681 cggatctcgc gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg 4741 ccctatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc 4801 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc 4861 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 4921 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 4981 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 5041 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 5101 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 5161 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 5221 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 5281 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 5341 ctgaaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 5401 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5461 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5521 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5581 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5641 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5701 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5761 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5821 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5881 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 5941 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 6001 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 6061 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 6121 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 6181 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 6241 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 6301 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 6361 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6421 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 6481 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6541 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6601 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6661 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6721 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6781 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6841 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6901 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6961 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7441 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7741 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7801 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7861 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7921 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7981 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8041 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8101 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8161 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8221 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8281 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8341 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8401 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8461 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8521 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8581 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8641 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8701 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8761 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8821 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8881 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8941 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 9001 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 9061 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 9121 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9181 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9241 tgccaagctg //