LOCUS V035289 9070 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035289 VERSION V035289 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3978 /label="Plat(NM_008872)" /note="Plat(NM_008872)" /gene="Plat" CDS 3979..3996 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 4040..4251 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4264..4532 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4559..5158 /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 5165..5753 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5827..6060 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6132..6253 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6272..6407 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6445..6461) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6469..6485 /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(6493..6523) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6538..6559 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6847..7435) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7606..8466) /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(8467..8571) /gene="bla" /label="AmpR promoter" primer_bind 9045..9061 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgaagaga gagctgctgt gtgtactgct gctttgtgga 2341 ctggctttcc cattgcctga ccagggaata catgggaggt tcagaagagg agcccggtcc 2401 tacagagcga cctgcagaga tgagccaacg cagacaactt accaacagca ccagtcgtgg 2461 ctccgaccca tgctcagaag cagccgggtg gaatattgcc ggtgcaacag cggcctggta 2521 caatgccact cagtgcctgt ccgaagttgc agcgaaccaa gatgcttcaa tggggggacg 2581 tgtcagcagg ccctgtattt ctctgacttt gtctgccagt gccctgatgg atttgtaggg 2641 aaacgctgtg acatagatac cagagcaaca tgctttgagg agcagggcat cacctacaga 2701 ggcacgtgga gcacagcaga aagtggggct gagtgcatca actggaatag cagtgttctg 2761 tcgctgaagc cctacaatgc aaggaggcca aatgccatca agctgggcct tgggaatcac 2821 aattactgca gaaacccaga ccgagacttg aagccctggt gctatgtctt taaggcaggg 2881 aagtatacca cggagttctg tagcacacca gcttgcccta agggaaaaag cgaggactgc 2941 tatgttggga aaggtgtgac ttaccgtggc acccacagcc tcaccacatc ccaggcctcc 3001 tgcctcccct ggaattccat agtcctcatg ggcaagagtt acacagcgtg gaggaccaac 3061 tcccaggcac tcggcctggg cagacacaat tattgtcgga atccagatgg tgatgccaga 3121 ccttggtgcc atgtgatgaa ggaccgaaag ctgacgtggg aatactgtga catgtcccca 3181 tgctccacct gtggcctgag gcagtacaaa cggcctcagt ttagaattaa aggaggactc 3241 tacacagaca tcacctcaca cccttggcag gctgccatct ttgtcaagaa caagaggtct 3301 cctggagaga gattcctttg tggaggggtg ctgatcagtt cctgctgggt gctgtcagct 3361 gcccactgct ttctagagag gttccccccc aatcatctta aagtggtctt gggcagaaca 3421 tacagggtgg tccccggaga ggaagaacag acatttgaga ttgaaaaata catagtccat 3481 gaggaatttg atgacgatac ttatgacaac gacatcgcat tactgcagct gaggtcacag 3541 tccaagcaat gtgcccaaga gagcagctct gttggcactg cctgcctccc tgaccccaac 3601 ctgcagctcc ctgactggac agagtgtgag ctttctggct acggcaagca tgaggcatcg 3661 tctccattct tctctgaccg gctgaaggag gctcacgtca gactgtaccc gtccagccgc 3721 tgcacctcac agcatctgtt taataaaacc gtcacgaaca acatgctgtg cgctggagac 3781 acccggagcg gaggcaacca agacctccac gatgcatgcc agggtgactc gggaggccct 3841 ctggtgtgca tgatcaataa acagatgaca ttgactggca ttatcagctg gggcctcggc 3901 tgtgggcaga aggatgtgcc tggggtctac acaaaggtta cgaattacct agactggatt 3961 cacgacaaca tgaagcaaca tcaccatcac catcactagc ggccgcgaag gatctgcgat 4021 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 4081 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 4141 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 4201 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4261 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4321 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4381 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4441 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4501 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat 4561 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 4621 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 4681 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4741 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4801 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4861 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4921 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4981 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 5041 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 5101 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 5161 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 5221 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5281 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5341 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5401 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5461 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5521 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5581 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5641 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5701 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 5761 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5821 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5881 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5941 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 6001 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 6061 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 6121 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 6181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6241 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 6301 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6361 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6421 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6481 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7261 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8581 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8641 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8701 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8761 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8821 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8881 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8941 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 9001 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9061 tgccaagctg //