LOCUS V029334 8851 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029334 VERSION V029334 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..2877 /label="Tnni1(NM_021467)" /note="Tnni1(NM_021467)" /gene="Tnni1" CDS 2884..2949 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2993..3204 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3217..3485 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3514..3523 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3544..4206 /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 4276..4329 /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 4330..4929 /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 4930..5518 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5592..5825 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5897..6018 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6058..6193 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6226..6242) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6250..6266 /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(6274..6304) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6319..6340 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6628..7216) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7387..8247) /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(8248..8352) /gene="bla" /label="AmpR promoter" primer_bind 8826..8842 /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 cggaagttga gaggaaatcc 2341 aagatcactg cctcccgtaa actcatgctg aagagcctga tgctagccaa ggccaaggag 2401 tgttgggagc aggaacacga ggagcgagag gctgagaagg tgcgttacct ctcggagcgc 2461 attcccacac tgcagacccg cggtctgtct ctcagtgccc ttcaggactt gtgccgagag 2521 ctccatgcca aggtagaggt ggtggatgag gagcgctatg atattgaagc caaatgcctc 2581 cacaacacca gagagatcaa ggacctgaag ctgaaggtgc tggacctccg tgggaagttc 2641 aagcgtcctc ccctgcgccg ggtccgtgtc tcagccgatg ccatgctccg agccctactg 2701 ggctctaagc acaaggtgtc catggatctg cgggccaacc tcaagtctgt gaagaaagaa 2761 gacacagaaa aggagcggcc agtcgaggta ggagactgga ggaagaatgt ggaggctatg 2821 tctggcatgg aaggccgcaa gaagatgttc gacgctgcta agtccccgac ctcccaggga 2881 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 2941 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 3001 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 3061 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 3121 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3181 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3241 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3301 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3361 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 3421 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 3481 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 3541 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 3601 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 3661 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 3721 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 3781 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 3841 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 3901 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 3961 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 4021 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 4081 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 4141 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 4201 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 4261 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4321 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4381 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 4441 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 4501 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 4561 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 4621 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 4681 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 4741 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 4801 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 4861 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 4921 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 4981 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5041 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5101 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5161 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5221 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5281 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5341 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5401 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5461 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5521 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 5581 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 5641 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 5701 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 5761 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 5821 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 5881 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5941 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 6001 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 6061 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 6121 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 6181 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 6241 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6301 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6361 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6421 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6481 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6541 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7141 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7201 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7261 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7321 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7381 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7441 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7501 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7561 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7621 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7681 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7741 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7801 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7861 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7921 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7981 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8041 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8101 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8161 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8221 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8281 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8341 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8401 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8461 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8521 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8581 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8641 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8701 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8761 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8821 acgttgtaaa acgacggcca gtgccaagct g //