LOCUS V020049 8170 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020049 VERSION V020049 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 1960..2171 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2184..2452 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 2492..2501 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2501..2566 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2582..3577 /label="Npdc1(NM_008721)" /note="Npdc1(NM_008721)" /gene="Npdc1" CDS 3593..3646 /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 3653..4252 /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 4265..4853 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4927..5160 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5232..5353 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5372..5507 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5545..5561) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5569..5585 /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(5593..5623) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5638..5659 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5947..6535) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6706..7566) /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(7567..7671) /gene="bla" /label="AmpR promoter" primer_bind 8145..8161 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca 1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc 2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta 2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct 2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc 2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc 2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt 2461 gaccggcgcc tactctagag ctagcgaatt cgccaccatg gactacaaag accatgacgg 2521 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagcttg gtaccctcga 2581 gatggcgacg cccgtccctc cgccttcccc gcggcacctg cggctgctac ggctgctgct 2641 ctccggcctc atcctgggag cggccctgaa tggtgccacc gcccgccgcc cggatgctac 2701 cacctgtcct ggaagcctgg attgtgcact gaagaggcgg gcaaaatgcc caccaggggc 2761 acatgcctgt ggaccatgcc ttcagtcctt ccaggaagac cagcgagggt tctgtgtgcc 2821 aaggaaacac ctatcttctg gggagggcct gccccagccc agactggaag aggagatcga 2881 ttctctggcc caggaactgg cactgaagga gaaggaggcg gggcactcaa ggctgaccgc 2941 acagcccttg cttgaggctg cgcagaagct cctggagcct gcagctaccc tgggattctc 3001 acagtggggt caacgactgg agcccgggct cccctctact catggaacct cttcacctat 3061 accccatact tccttgagtt cccgagcatc ctctggacct gtgcagatgt ccccgctgga 3121 gccccaggga aggcatggga atggcctcac ccttgtgctg atcctggctt tctgcttagc 3181 aagctcagcc gccctggctg tggcagccct ctgctggtgt aggcttcagc gagagatccg 3241 cctgactcag aaggctgact atgctgccac agctaagggg cccacttcac cttcaacacc 3301 acgaatctcg cctggggatc agcggctggc acacagtgct gagatgtatc attatcagca 3361 ccagagacag cagatgctgt gcctggagcg gcataaggaa cctccaaagg aactagagtc 3421 agcttcctcc gatgaggaaa atgaagatgg cgacttcact gtgtatgaat gcccaggcct 3481 ggcgccaaca ggggagatgg aggtgcgcaa cccactgttt gaccattcca cgctgtcagc 3541 accagtgcct ggtccccatt ccttgccgcc gctgcaggga tccgcggccg ctgagggcag 3601 aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggcccttccg ggatgaccga 3661 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 3721 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 3781 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 3841 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 3901 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 3961 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4021 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4081 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4141 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4201 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaatctaggt 4261 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 4321 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 4381 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 4441 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 4501 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 4561 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 4621 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 4681 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 4741 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 4801 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 4861 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 4921 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 4981 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5041 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5101 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5161 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5221 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5281 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5341 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 5401 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 5461 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 5521 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 5581 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 5641 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 5701 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 5761 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 5821 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 5881 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 5941 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6001 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6061 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6121 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6181 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6241 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6301 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6361 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 6421 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 6481 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 6541 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 6601 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 6661 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 6721 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 6781 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 6841 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 6901 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 6961 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7021 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7081 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7141 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7201 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7261 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7321 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 7381 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7441 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 7501 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 7561 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 7621 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 7681 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 7741 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 7801 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 7861 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 7921 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 7981 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8041 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8101 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8161 tgccaagctg //