LOCUS V047328 9946 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047328 VERSION V047328 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..3972 /label="GNS(NM_002076)" /note="GNS(NM_002076)" /gene="GNS" CDS 3979..4044 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4088..4299 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4312..4580 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4609..4618 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4639..5301 /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 5371..5424 /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 5425..6024 /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 6025..6613 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6687..6920 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6992..7113 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7153..7288 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7321..7337) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7345..7361 /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(7369..7399) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7414..7435 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7723..8311) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8482..9342) /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(9343..9447) /gene="bla" /label="AmpR promoter" primer_bind 9921..9937 /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 gccaccatgc ggctcctgcc tctagcccca 2341 ggtcggctcc ggcggggcag cccccgccac ctgccctcct gcagcccagc gctgctactg 2401 ctggtgctgg gcggctgcct gggggtcttc ggggtggctg cgggaacccg gaggcccaac 2461 gtggtgctgc tcctcacgga cgaccaggac gaagtgctcg gcggcatgac accgctaaag 2521 aaaaccaaag ctctcatcgg agagatgggg atgacttttt ccagtgctta tgtgccaagt 2581 gctctctgct gccccagcag agccagtatc ctgacaggaa agtacccaca taatcatcac 2641 gttgtgaaca acactctgga ggggaactgc agtagtaagt cctggcagaa gatccaagaa 2701 ccaaatactt tcccagcaat tctcagatca atgtgtggtt atcagacctt ttttgcaggg 2761 aaatatttaa atgagtacgg agccccagat gcaggtggac tagaacacgt tcctctgggt 2821 tggagttact ggtatgcctt ggaaaagaat tctaagtatt ataattacac cctgtctatc 2881 aatgggaagg cacggaagca tggtgaaaac tatagtgtgg actacctgac agatgttttg 2941 gctaatgtct ccttggactt tctggactac aagtccaact ttgagccctt cttcatgatg 3001 atcgccactc cagcgcctca ttcgccttgg acagctgcac ctcagtacca gaaggctttc 3061 cagaatgtct ttgcaccaag aaacaagaac ttcaacatcc atggaacgaa caagcactgg 3121 ttaattaggc aagccaagac tccaatgact aattcttcaa tacagttttt agataatgca 3181 tttaggaaaa ggtggcaaac tctcctctca gttgatgacc ttgtggagaa actggtcaag 3241 aggctggagt tcactgggga gctcaacaac acttacatct tctatacctc agacaatggc 3301 tatcacacag gacagttttc cttgccaata gacaagagac agctgtatga gtttgatatc 3361 aaagttccac tgttggttcg aggacctggg atcaaaccaa atcagacaag caagatgctg 3421 gttgccaaca ttgacttggg tcctactatt ttggacattg ctggctacga cctaaataag 3481 acacagatgg atgggatgtc cttattgccc attttgagag gtgccagtaa cttgacctgg 3541 cgatcagatg tcctggtgga ataccaagga gaaggccgta acgtcactga cccaacatgc 3601 ccttccctga gtcctggcgt atctcaatgc ttcccagact gtgtatgtga agatgcttat 3661 aacaatacct atgcctgtgt gaggacaatg tcagcattgt ggaatttgca gtattgcgag 3721 tttgatgacc aggaggtgtt tgtagaagtc tataatctga ctgcagaccc agaccagatc 3781 actaacattg ctaaaaccat agacccagag cttttaggaa agatgaacta tcggttaatg 3841 atgttacagt cctgttctgg gccaacctgt cgcactccag gggtttttga ccccggatac 3901 aggtttgacc cccgtctcat gttcagcaat cgcggcagtg tcaggactcg aagattttcc 3961 aaacatcttc tgggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 4021 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 4081 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 4141 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 4201 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 4261 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 4321 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 4381 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4441 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4501 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4561 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4621 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4681 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4741 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4801 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4861 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4921 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4981 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 5041 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 5101 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 5161 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 5221 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 5281 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 5341 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 5401 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 5461 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5521 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5581 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5641 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5701 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5761 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5821 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5881 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5941 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 6001 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 6061 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 6121 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 6181 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 6241 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 6301 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 6361 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 6421 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6481 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6541 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6601 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6661 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6721 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6781 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6841 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6901 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6961 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 7021 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 7081 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 7141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 7201 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 7261 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 7321 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7381 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7441 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7501 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7561 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7621 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7681 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7741 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7801 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7861 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7921 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7981 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 8041 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 8101 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 8161 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 8221 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 8281 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 8341 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8401 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8461 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8521 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8581 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8641 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8701 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8761 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8821 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8881 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8941 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 9001 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 9061 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 9121 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 9181 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 9241 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 9301 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9361 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9421 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9481 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9541 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9601 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9661 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9721 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9781 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9841 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9901 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //