LOCUS V029046 9613 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029046 VERSION V029046 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..3639 /label="Ifrd2(NM_025903)" /note="Ifrd2(NM_025903)" /gene="Ifrd2" CDS 3646..3711 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3755..3966 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3979..4247 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4276..4285 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4306..4968 /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 5038..5091 /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 5092..5691 /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 5692..6280 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6354..6587 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6659..6780 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6820..6955 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6988..7004) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7012..7028 /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(7036..7066) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7081..7102 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7390..7978) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8149..9009) /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(9010..9114) /gene="bla" /label="AmpR promoter" primer_bind 9588..9604 /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 ctcgtgctcg gaagggcaat 2341 gcgctccgca agggcggtca gcgccgtgga ggaggtgcca ggagcagtac ccaggctgac 2401 tctggctcca gtgaggatga agcggccagt gaggcccgga gcaccaccag tgactgtccc 2461 agccttctca gcgccaccac agaggactgc ctgggtggcg aagcggtgga cgaacagagc 2521 cagcaggaaa acctggagga gaagctgaaa ggttatgtgg actgcctgac tgacaagagt 2581 gccaagaccc gtcaaggagc cctggagagc ctgcgcctgg ccctggcctc ccgcctactt 2641 ccggactttc tgctggagcg cagcctcact ctggcggatg ccctggaaaa gtgccttaag 2701 aaagggaagg gtgaagaaca ggctctggct gctgctgtgc taggcattct ctgtgtgcag 2761 ttgggccctg gacccaaggg cgaggagctg ttccgtagcc tgcagccctt gctcatctcg 2821 gtgctcagtg acagtacagc aagccccact gcccggctcc attgtgcttc tgctcttggc 2881 ttgggttgct atgtggctgc caccgacgtc caggacctgg tctcttgctt ggcctgcttg 2941 gaaggtgttt tcagctggtc ctgtggcact agtggctctg cggcgtctct ggttcccgcc 3001 agcctccatg gccttctctg tgctgccctg caggcctggg cattgctgct caccatctgt 3061 cctggtaccc acatcagcca tatccttgac aggcagctgc cccggctgcc ccagctcttg 3121 tccagtgaaa gtgtgaacct gcggattgct gctggtgaag ccatcgcgct actctttgag 3181 cttgcccggg accttgagga ggacttcgtc tatgaagaca tggaggccct ctgtggatct 3241 ctgcgtaccc tagccaccga cagcaataag taccgtgcca aggttgaccg gcggcgccag 3301 cgctccattt tccgagctgt gctgcacttt gttgagggtg gtgaatgtga ggaggagaca 3361 gtccgcttcg gactggaagt gctctacata gacagctggg ctcgccaccg catctacaca 3421 gccttcaaag atgtgttggg ctctggcata cactatcacc tccagaacaa tgagcttctc 3481 cgtgacatct ttggcctggg ccccgtgcta gtgttggatg ctgctgccct gaaggcctgc 3541 aagatttcac gctttgaaaa gcacctgtac aatgctgcag ccttcaaagc ccggaccaag 3601 gcccgtagtc gtgcaaggga caagcgggca gatatcttgg gatccgacta caaagaccat 3661 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3721 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3781 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3841 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3901 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3961 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4021 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4081 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4141 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4201 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4261 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4321 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4381 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4441 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4501 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4561 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4621 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4681 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4741 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4801 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4861 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 4921 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4981 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 5041 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 5101 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5161 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5221 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5281 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5341 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5401 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5461 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5521 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5581 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5641 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5701 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5761 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5821 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5881 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5941 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6001 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6061 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6121 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6181 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6241 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6301 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6361 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6421 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6481 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6541 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6601 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6661 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6721 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6781 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6841 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6901 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6961 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 7021 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7081 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7141 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7201 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7561 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7801 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8161 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8221 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8281 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8341 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8401 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8461 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8521 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8581 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8641 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8701 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8761 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8821 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8881 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8941 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9001 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9061 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9121 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9181 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9241 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9301 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9361 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9421 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9481 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9541 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9601 cagtgccaag ctg //