LOCUS V034675 8887 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034675 VERSION V034675 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 2303..2941 /label="VHL(NM_000551)" /note="VHL(NM_000551)" /gene="VHL" CDS 2948..2974 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2978..3004 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3008..3034 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3085..3296 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3309..3577 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3612..4319 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 4329..4382 /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 4383..4982 /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 4983..5570 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5644..5877 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5949..6070 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6089..6224 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6262..6278) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6286..6302 /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(6310..6340) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6355..6376 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6664..7252) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7423..8283) /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(8284..8388) /gene="bla" /label="AmpR promoter" primer_bind 8862..8878 /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 gaattcgcca ccatgccccg gagggcggag aactgggacg aggccgaggt 2341 aggcgcggag gaggcaggcg tcgaagagta cggccctgaa gaagacggcg gggaggagtc 2401 gggcgccgag gagtccggcc cggaagagtc cggcccggag gaactgggcg ccgaggagga 2461 gatggaggcc gggcggccgc ggcccgtgct gcgctcggtg aactcgcgcg agccctccca 2521 ggtcatcttc tgcaatcgca gtccgcgcgt cgtgctgccc gtatggctca acttcgacgg 2581 cgagccgcag ccctacccaa cgctgccgcc tggcacgggc cgccgcatcc acagctaccg 2641 aggtcacctt tggctcttca gagatgcagg gacacacgat gggcttctgg ttaaccaaac 2701 tgaattattt gtgccatctc tcaatgttga cggacagcct atttttgcca atatcacact 2761 gccagtgtat actctgaaag agcgatgcct ccaggttgtc cggagcctag tcaagcctga 2821 gaattacagg agactggaca tcgtcaggtc gctctacgaa gatctggaag accacccaaa 2881 tgtgcagaaa gacctggagc ggctgacaca ggagcgcatt gcacatcaac ggatgggaga 2941 tctcgagtac ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta 3001 tgctagctat ccttatgatg tcccagacta cgcataggga tccgcggccg cgaaggatct 3061 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 3121 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg 3181 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 3241 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga 3301 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc 3361 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg 3421 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga 3481 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg 3541 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc 3601 tagacgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca 3661 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 3721 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg 3781 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct 3841 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca 3901 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 3961 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg 4021 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc 4081 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc 4141 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg 4201 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg 4261 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg 4321 gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc 4381 ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg 4441 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg 4501 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 4561 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 4621 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 4681 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 4741 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 4801 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 4861 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 4921 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 4981 gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5041 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5101 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5161 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5221 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5281 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 5341 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 5401 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 5461 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 5521 tccgccttcg ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt 5581 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5641 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5701 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5761 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5821 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5881 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 5941 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6001 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6061 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6121 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6181 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6241 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7081 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7321 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7381 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7441 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7501 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7561 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7621 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7681 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7741 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7801 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7861 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7921 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7981 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8041 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8101 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8161 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8221 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8281 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8401 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8461 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8521 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8581 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8641 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8701 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8761 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8821 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8881 caagctg //