LOCUS V029357 8671 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029357 VERSION V029357 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..2697 /label="FABP1(NM_001443)" /note="FABP1(NM_001443)" /gene="FABP1" CDS 2704..2769 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2813..3024 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3037..3305 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3334..3343 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3364..4026 /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 4096..4149 /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 4150..4749 /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 4750..5338 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5412..5645 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5717..5838 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5878..6013 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6046..6062) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6070..6086 /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(6094..6124) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6139..6160 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6448..7036) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7207..8067) /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(8068..8172) /gene="bla" /label="AmpR promoter" primer_bind 8646..8662 /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 gccaccatga gtttctccgg caagtaccaa 2341 ctgcagagcc aggaaaactt tgaagccttc atgaaggcaa tcggtctgcc ggaagagctc 2401 atccagaagg ggaaggatat caagggggtg tcggaaatcg tgcagaatgg gaagcacttc 2461 aagttcacca tcaccgctgg gtccaaagtg atccaaaacg aattcacggt gggggaggaa 2521 tgtgagctgg agacaatgac aggggagaaa gtcaagacag tggttcagtt ggaaggtgac 2581 aataaactgg tgacaacttt caaaaacatc aagtctgtga ccgaactcaa cggcgacata 2641 atcaccaata ccatgacatt gggtgacatt gtcttcaaga gaatcagcaa gagaattgga 2701 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 2761 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 2821 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 2881 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 2941 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3001 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3061 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3121 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3181 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 3241 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 3301 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 3361 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 3421 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 3481 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 3541 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 3601 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 3661 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 3721 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 3781 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 3841 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 3901 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 3961 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 4021 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 4081 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4141 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4201 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 4261 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 4321 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 4381 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 4441 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 4501 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 4561 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 4621 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 4681 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 4741 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 4801 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 4861 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 4921 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 4981 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5041 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5101 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5161 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5221 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5281 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5341 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 5401 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 5461 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 5521 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 5581 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 5641 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 5701 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5761 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 5821 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 5881 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 5941 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 6001 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 6061 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6121 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6181 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6241 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6301 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6361 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 7981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8221 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8281 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8341 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8401 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8461 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8521 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8581 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8641 acgttgtaaa acgacggcca gtgccaagct g //