LOCUS V030611 8767 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030611 VERSION V030611 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..3597 /label="RNF135(NM_032322)" /note="RNF135(NM_032322)" /gene="RNF135" CDS 3604..3693 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" promoter 3737..3948 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3961..4229 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4256..4855 /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 4862..5450 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5524..5757 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5829..5950 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5969..6104 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6142..6158) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6166..6182 /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(6190..6220) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6235..6256 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6544..7132) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7303..8163) /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(8164..8268) /gene="bla" /label="AmpR promoter" primer_bind 8742..8758 /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 catggcgggc ctgggcctgg gctccgccgt tcccgtgtgg 2341 ctggccgagg acgacctcgg ctgcatcatc tgccaggggc tgctggactg gcccgccacg 2401 ctgccctgcg gccacagctt ctgccgccac tgcctggagg ccctgtgggg cgcccgcgac 2461 gcccgccgct gggcctgccc cacttgccgc cagggcgccg cgcagcagcc gcacctgcgg 2521 aagaacacgc tactgcagga cctggccgac aagtaccgcc gcgccgcacg cgagatacag 2581 gcgggctccg accctgccca ctgcccctgc ccgggctcca gttccctctc cagcgcggcc 2641 gcgaggcccc ggcgccgccc ggaactgcag cgggtggcag tagagaagag catcacagaa 2701 gttgctcagg agctgacaga gctggtggaa catcttgtag acattgtcag aagcctgcag 2761 aatcagaggc ccctatcaga atctggacca gacaacgaac tgagcatcct gggcaaggct 2821 ttttcttctg gggtggatct ttccatggct tctccaaagc tggtgacttc cgacacagct 2881 gcagggaaaa tcagagatat tctccatgac ctagaagaaa ttcaggaaaa attacaagaa 2941 agcgtcacct ggaaagaggc tcctgaagca caaatgcagg gagaactcct ggaagccccg 3001 tcttcctcct catgcccatt gcctgaccag agccaccctg cactcaggag agcttctcgg 3061 tttgctcagt gggccatcca tccaaccttt aacttgaaga gcctttcctg cagcctggag 3121 gtgtccaagg attcccgtac agtgactgtg tctcaccgcc cacaacccta tcgctggagc 3181 tgtgagaggt tttctaccag ccaggtctta tgttcccagg ccctgtcttc tggaaagcat 3241 tactgggaag tggacactag gaattgcagc cactgggcag ttggggtggc ttcctgggag 3301 atgagccgcg accaggtcct gggaaggact atggactctt gttgtgtgga atggaagggg 3361 actagccagc tctctgcatg gcacatggtc aaggaaactg tccttggctc agacagacct 3421 ggggtggtgg gcatctggct gaaccttgag gagggaaagc ttgccttcta ttcagtggac 3481 aatcaggaga agcttctgta tgagtgtacc atctctgcct cctctccttt gtaccctgcc 3541 ttctggctgt atggcttaca tcctggaaat tacctgataa taaagcaagt aaaggtggga 3601 tccgaacaaa aactaatatc agaggaagat ttggaacaga aactaatcag tgaagaagat 3661 ttagagcaaa agttaatttc cgaggaggac ttatagcggc cgcgaaggat ctgcgatcgc 3721 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 3781 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 3841 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 3901 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 3961 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4021 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4081 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4141 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 4201 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 4261 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 4321 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 4381 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4441 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4501 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4561 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4621 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4681 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4741 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4801 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 4861 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4921 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4981 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5041 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5101 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5161 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5221 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 5281 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5341 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5401 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 5461 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5521 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5581 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5641 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5701 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5761 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 5821 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5881 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5941 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6001 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6061 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6121 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6181 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6241 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6301 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6961 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7261 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7321 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7381 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7441 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7501 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7561 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7621 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7681 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7741 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7801 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7861 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7921 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7981 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8041 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8101 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8161 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8221 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8281 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8341 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8401 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8461 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8521 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8581 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8641 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8701 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8761 caagctg //