LOCUS V039739 9205 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039739 VERSION V039739 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..3231 /label="DNASE1L3(NM_004944)" /note="DNASE1L3(NM_004944)" /gene="DNASE1L3" CDS 3238..3303 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3347..3558 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3571..3839 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3868..3877 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3898..4560 /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 4630..4683 /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 4684..5283 /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 5284..5872 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5946..6179 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6251..6372 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6412..6547 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6580..6596) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6604..6620 /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(6628..6658) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6673..6694 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6982..7570) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7741..8601) /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(8602..8706) /gene="bla" /label="AmpR promoter" primer_bind 9180..9196 /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 gccaccatgt cacgggagct ggccccactg 2341 ctgcttctcc tcctctccat ccacagcgcc ctggccatga ggatctgctc cttcaacgtc 2401 aggtcctttg gggaaagcaa gcaggaagac aagaatgcca tggatgtcat tgtgaaggtc 2461 atcaaacgct gtgacatcat actcgtgatg gaaatcaagg acagcaacaa caggatctgc 2521 cccatactga tggagaagct gaacagaaat tcaaggagag gcataacgta caactatgtg 2581 attagctctc ggcttggaag aaacacatat aaagaacaat atgcctttct ctacaaggaa 2641 aagctggtgt ctgtgaagag gagttatcac taccatgact atcaggatgg agacgcagat 2701 gtgttttcca gggagccctt tgtggtctgg ttccaatctc cccacactgc tgtcaaagac 2761 ttcgtgatta tccccctgca caccacccca gagacatccg ttaaggagat cgatgagttg 2821 gttgaggtct acacggacgt gaaacaccgc tggaaggcgg agaatttcat tttcatgggt 2881 gacttcaatg ccggctgcag ctacgtcccc aagaaggcct ggaagaacat ccgcttgagg 2941 actgacccca ggtttgtttg gctgatcggg gaccaagagg acaccacggt gaagaagagc 3001 accaactgtg catatgacag gattgtgctt agaggacaag aaatcgtcag ttctgttgtt 3061 cccaagtcaa acagtgtttt tgacttccag aaagcttaca agctgactga agaggaggcc 3121 ctggatgtca gcgaccactt tccagttgaa tttaaactac agtcttcaag ggccttcacc 3181 aacagcaaaa aatctgtcac tctaaggaag aaaacaaaga gcaaacgctc cggatccgac 3241 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 3301 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca 3361 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga 3421 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag 3481 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg 3541 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc 3601 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt 3661 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc 3721 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg 3781 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc 3841 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc 3901 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg 3961 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa 4021 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac 4081 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc 4141 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc 4201 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc 4261 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg 4321 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac 4381 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc 4441 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc 4501 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc 4561 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc 4621 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 4681 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 4741 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 4801 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 4861 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 4921 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 4981 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 5041 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 5101 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 5161 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 5221 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 5281 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5341 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5401 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5461 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5521 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 5581 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 5641 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg 5701 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 5761 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 5821 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc 5881 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 5941 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg 6001 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6061 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6121 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6181 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 6241 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 6301 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6361 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc 6421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 6481 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 6541 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg 6601 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6661 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 6721 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 6781 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 6841 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 6961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7381 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 7441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7801 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7861 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7921 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7981 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8041 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8101 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8161 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8221 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8281 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8341 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8401 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8461 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8521 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8581 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8641 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 8701 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 8761 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 8821 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 8881 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 8941 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 9001 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 9061 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 9121 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 9181 taaaacgacg gccagtgcca agctg //