LOCUS V046073 8584 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046073 VERSION V046073 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..2610 /label="COMMD6(NM_203497)" /note="COMMD6(NM_203497)" /gene="COMMD6" CDS 2617..2682 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2726..2937 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2950..3218 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3247..3256 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3277..3939 /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 4009..4062 /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 4063..4662 /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 4663..5251 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5325..5558 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5630..5751 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5791..5926 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5959..5975) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5983..5999 /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(6007..6037) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6052..6073 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6361..6949) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7120..7980) /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(7981..8085) /gene="bla" /label="AmpR promoter" primer_bind 8559..8575 /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 aggcgtccag cgagccgccg 2341 ctggatgcta agtccgatgt caccaaccag cttgtagatt ttcagtggaa actgggtatg 2401 gctgtgagct cagacacttg cagatctctt aagtatcctt acgttgcagt gatgctaaaa 2461 gtggcagatc attcaggcca agtaaagacc aagtgctttg aaatgacgat tccacagttt 2521 cagcctcagc tcctggcaac ctccagtctc ctttctgctt ctaatttcta cagacagttc 2581 aaggaaattg ctgcagttat tgaaacggtg ggatccgact acaaagacca tgacggtgat 2641 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 2701 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 2761 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 2821 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 2881 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 2941 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3001 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3061 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3121 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 3181 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 3241 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 3301 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 3361 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 3421 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 3481 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 3541 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 3601 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 3661 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 3721 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 3781 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 3841 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 3901 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 3961 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4021 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4081 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4141 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4201 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4261 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4321 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 4381 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 4441 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 4501 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 4561 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 4621 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 4681 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 4741 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 4801 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 4861 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 4921 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 4981 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5041 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 5101 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 5161 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 5221 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 5281 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 5341 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 5401 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 5461 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 5521 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 5581 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 5641 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5701 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5761 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 5821 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 5881 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 5941 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6001 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6061 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6121 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6181 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6241 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6301 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6361 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6421 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6481 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6541 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6601 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6661 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6721 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6781 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6841 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6901 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6961 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7021 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7081 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7141 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7201 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7261 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7321 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7381 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7441 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7501 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7561 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7621 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7681 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7741 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7801 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7861 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7921 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7981 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8041 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8101 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8161 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8221 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8281 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8341 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8401 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8461 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8521 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8581 gctg //