LOCUS V028797 10036 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028797 VERSION V028797 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..4062 /label="Nudcd1(NM_026149)" /note="Nudcd1(NM_026149)" /gene="Nudcd1" CDS 4069..4134 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4178..4389 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4402..4670 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4699..4708 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4729..5391 /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 5461..5514 /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 5515..6114 /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 6115..6703 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6777..7010 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7082..7203 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7243..7378 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7411..7427) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7435..7451 /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(7459..7489) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7504..7525 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7813..8401) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8572..9432) /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(9433..9537) /gene="bla" /label="AmpR promoter" primer_bind 10011..10027 /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 aggcggccaa ttgctctctg 2341 cgggtgaaaa gacctctgct ggatccccgc ttcgagggct acaagctgtc tctcgaaccg 2401 ctgccctgct accagctgga gcttgatgca gctgtggcag aagtaaaact tcgagatgac 2461 cagtatacat tagaacacat gcatgctttt gggatgtata attatctgca ctgtgatgcc 2521 tggtatcaag acagtgtcta ctacattgat aaccttggaa gaattatgaa cttaacagtg 2581 atgctggaca ccgccttagg aaaaccccga gaggtgttcc gccttcctac agatgtgaca 2641 gcatatgaca atcgtctttg tgcatctatc catttcacat ccccttcctg ggttaccttg 2701 tcagatggaa ctggaagact gtatgtcatt ggaacaggtg accgtggaaa cagtcctgaa 2761 aaatgggaga ttatgtttaa tgaagagctt gggaaccctt ttattataat tcacagtatc 2821 tcactgctga aagctgaaga acattcagtg gctatcctgc ttcttcgaat agagaaggag 2881 gaattggacc tgaaaggaag tggtttctat gtttctttgg agtgggtcgt tgtcactaag 2941 aagaaagaag atagtaaaaa gtatgaaatt acaaagcatc atgttctccg tggaaagtca 3001 gtgccacatt atgctgcaat tgaacctaat ggaaatggtc taatgattgt atcttacaag 3061 tccttcaagg ttatttcggg agatcaggat cttgaagaga atatggatga agacaggcca 3121 gagaaaatca aagtagaacc tctgtattac tggcaacaaa gtgaggatga tttgacagta 3181 acggtcaggc ttccagaaag cagtactaag gaagatattc aaatacaatt tttgcctgat 3241 aatatcaaca ttaagctgaa ggatatccaa gttttggaag gaaaactcta ttcatctatt 3301 gatcacgaag gcagcacatg gacaattaaa gagaatgata gcttggagat ttctttgatt 3361 aagaagaatg aaggactgat gtggcctgag ctggtggttg gtgataaaca aggtgaactt 3421 ttaagagacc ctgcccaatg tgctgcaata gcagaacgct tgatgcacct gacctctgat 3481 gaactgaatc caaatccaga taaggaaaag cctgcatgta atgctcaaga attagaagaa 3541 tgtgatattt tctttgaaga gagctccagt ttatgcagat ttgatggcaa tacattacaa 3601 acaactcatg tggtgaatct tggcagcaac cagtacctgt tctcagtcat tgtggatcct 3661 aaagaaatgc cgtgcttctg tctacgccat gatgtcgatg ctctcctctg gcaaccacac 3721 tgcagcaagc aggacgacat gtgggagcac attgccacct tcaatgccct aggttacgtc 3781 caagcctcaa agagagacaa gaaatttttt gcatgtgctc caaactactc gtatgcagcc 3841 ctttgtgagt gccttcgccg agtattcatc tatcggcagc ccacccccat gtccactgtg 3901 ctgtacaaca ggaaggaagg caggcatgtg gggcaagttg ctaagcagca ggtagcaagc 3961 ctagaaacca atgatcctat tttaggcttt caggcaacaa atgagagatt atttgttctc 4021 actaccaaaa acctcttttt aataagagta aatacggaga atggatccga ctacaaagac 4081 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 4141 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 4201 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 4261 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 4321 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 4381 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 4441 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 4501 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 4561 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 4621 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 4681 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 4741 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 4801 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 4861 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 4921 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 4981 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 5041 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 5101 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 5161 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 5221 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 5281 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 5341 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 5401 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 5461 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 5521 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 5581 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 5641 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5701 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5761 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5821 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5881 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5941 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 6001 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 6061 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 6121 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 6181 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 6241 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 6301 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 6361 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 6421 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 6481 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 6541 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 6601 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 6661 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 6721 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 6781 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 6841 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6901 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6961 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 7021 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 7081 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 7141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 7201 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 7261 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 7321 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 7381 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 7441 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 7501 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 7561 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 7621 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 7681 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 7741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 7801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 8041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 8101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 8161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 8221 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 8281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 8341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 8401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 8461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 8521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 8581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 8641 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 8701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 8761 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 8821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 8881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 9001 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 9061 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 9121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 9181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 9241 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 9301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 9361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 9421 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 9481 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 9541 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 9601 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 9661 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 9721 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 9781 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 9841 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9901 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9961 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 10021 ggccagtgcc aagctg //