LOCUS V036367 10024 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036367 VERSION V036367 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..4050 /label="Ttc39a(NM_001145948)" /note="Ttc39a(NM_001145948)" /gene="Ttc39a" CDS 4057..4122 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4166..4377 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4390..4658 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4687..4696 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4717..5379 /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 5449..5502 /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 5503..6102 /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 6103..6691 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6765..6998 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7070..7191 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7231..7366 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7399..7415) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7423..7439 /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(7447..7477) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7492..7513 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7801..8389) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8560..9420) /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(9421..9525) /gene="bla" /label="AmpR promoter" primer_bind 9999..10015 /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 gccaccatgg gtcagaaggg ccacaaggac 2341 tccctttatg attgtaaagg gtctcctgtg agcagcctcc atgaggccct ggaccagtgc 2401 atgacagccc tggacctctt ccttaccaac cagttctcag aagcactcag ctacctcaaa 2461 cccagaacca aggagagcat gtatcactcc ctgacgtatg ccaccatcct ggagatgcag 2521 gccatgatga ccttcgaccc ccaggacatc ctgcttgctg gcaacatgat gaaagaagcg 2581 cagtcgctgt gtcagaggca tcgaaggaag tcttctatga cggactcatt cagcaacctg 2641 gtgcaccgtc ctaccattga ccagttcaca gaagaggaaa tccatgcaga ggtctgttat 2701 gcagaatgcc tactacagag agcggccctg accttcctgc aggacgagaa catggtgagc 2761 ttcatcaagg gcggcatcaa agtgcggaac agctaccaga cttacaagga actggatagc 2821 ctcgtccagt cctcgcaata ctctaaggga gagagccaca gacactttga aggaggggtg 2881 aagcttggcg taggagcctt caacctgacg ctgtccatgc ttcccaccag gattctgagg 2941 ctgctggagt ttgttgggtt ttcaggaaac aaggactatg ggctgctgca gctggaggag 3001 ggcgcaaccg gacacagctt ccgtgcagtg ctgtgcgtca tgctgttgct gtgttatcac 3061 accttcctca ccttcgtgct cggtactggg aatgtcaaca tcgaggaggc agagaagctg 3121 ctgaagccct acctgaaccg ataccccaag ggcgccatct tcctgttctt tgctgggcgg 3181 attgaagcta tcaaaggaaa tattgatgct gctgtccggc ggtttgagga gtgctgcgag 3241 gcccagcagc actggaagca gtttcaccac atgtgctact gggagctgat gtggtgcttt 3301 acctacaagg gccagtggaa aatggcctac ttctacgccg acctgctgag caaggagaac 3361 tcctggtcca aggccaccta catctacatg aaggctgcct acctcagcat gtttgggaag 3421 gaagactaca agccattcgg ggacaatgaa gtggagttat tcagggccgt gccaggcttg 3481 aagctcaaga ttgctgggaa atctctaccc acagagaagt ttgctatccg gaagtccagg 3541 cgctacctct cccccaaccc catctccttg ccgatccctg ctctggaaat gatgtacatc 3601 tggaatggct atgctgtgat tgggaagcag ccgacactca cggatgggat gctggaggtc 3661 atcaccaagg ctgaagagat gctggcaatg ggcccagaga atgagtactc agcagatgac 3721 gactgcttgg ttaaattgct gaaaggcctg tgtctgaaat acctgggccg aatccaggag 3781 gcagaggaga atttcaggag catttctgcc aatgaaaaga agattaaata tgaccattac 3841 ttgattccaa atgccctact ggaactggcc ctgctattta tggagcaagg cagaaatgag 3901 gaggccatca aactcttaga gtctgcaaag caaaactaca agaactactc catggagtca 3961 aggacacatt ttcgaatcca ggcagccaca ctccaggcca gatcttccct agaagatggc 4021 aacagatctc tagtctcgtc agtgtccttg ggatccgact acaaagacca tgacggtgat 4081 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 4141 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 4201 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 4261 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 4321 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 4381 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 4441 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4501 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4561 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4621 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4681 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4741 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4801 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4861 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4921 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4981 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 5041 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 5101 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 5161 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 5221 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 5281 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 5341 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 5401 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 5461 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 5521 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5581 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5641 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5701 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5761 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5821 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5881 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5941 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 6001 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 6061 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 6121 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 6181 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 6241 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 6301 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6361 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6421 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6481 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6541 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6601 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6661 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6721 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6781 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6841 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6901 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6961 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 7021 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 7081 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 7141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 7201 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 7261 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 7321 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 7381 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7441 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7501 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7621 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7681 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7741 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 8041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 8101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 8161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 8221 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 8281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 9001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 9061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 9121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 9181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 9241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9541 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9601 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9661 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9721 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9781 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9841 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9901 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9961 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 10021 gctg //