LOCUS V028861 9127 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028861 VERSION V028861 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..3981 /label="CBX4(NM_003655)" /note="CBX4(NM_003655)" /gene="CBX4" CDS 3988..4053 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4097..4308 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4321..4589 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4616..5215 /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 5222..5810 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5884..6117 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6189..6310 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6329..6464 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6502..6518) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6526..6542 /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(6550..6580) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6595..6616 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6904..7492) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7663..8523) /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(8524..8628) /gene="bla" /label="AmpR promoter" primer_bind 9102..9118 /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 catggagctg ccagctgttg gcgagcacgt cttcgcggtg 2341 gagagcatcg agaagaagcg gatccgcaag ggcagagtgg agtatctggt gaaatggaga 2401 ggctggtcgc ccaaatataa cacgtgggaa ccggaggaga acatcctgga ccccaggctg 2461 ctgatcgcct tccagaacag ggaacggcag gagcagctga tgggatatcg gaagagaggg 2521 ccgaagccca aaccgctagt ggtgcaggtg cctacctttg cccgtcgttc caatgtcctg 2581 accggcctcc aggactcctc cactgacaac cgtgccaagc tggatttggg cgcgcagggg 2641 aagggccagg ggcatcagta cgagctcaac agcaagaagc accaccagta ccagccgcac 2701 agcaaggagc gggcgggcaa gcccccgccg ccgggcaaga gcggcaagta ctactaccag 2761 ctcaacagca agaagcacca cccctaccag cccgacccca aaatgtacga cctgcagtac 2821 cagggcggcc acaaggaggc gcccagcccc acctgcccgg acctgggggc caagagccac 2881 ccgcccgaca agtgggcgca aggtgcgggg gccaaaggct acctgggggc ggtgaagccc 2941 ctggccggtg cggcgggtgc tccaggcaaa ggctccgaga agggcccccc caacggaatg 3001 atgccggccc ccaaagaggc tgtgacgggc aacgggattg ggggcaagat gaagatagtc 3061 aagaacaaga acaagaacgg acgcatcgtg atcgtgatga gcaaatacat ggagaacggc 3121 atgcaggcgg tgaagatcaa gtccggcgag gtggcagagg gggaggctcg ctcccccagc 3181 cacaagaagc gggcagccga cgagcgccac cctcctgccg acaggacttt taaaaaggcg 3241 gcgggcgcag aggagaagaa ggtggaggcg ccgcccaaga ggagggagga ggaggtgtcc 3301 ggggttagcg atccgcagcc ccaggatgcc ggctcccgca agctgtcccc gaccaaggag 3361 gcctttggag agcagcccct gcagctcacc accaagcccg acctgcttgc ctgggacccg 3421 gcccggaaca cgcacccgcc ctcacaccac ccgcacccgc acccccatca ccaccaccac 3481 caccaccacc accaccacca cgccgtcggc ctgaatctct cccacgtgcg caagcgctgc 3541 ctctccgaga cccacggcga gcgcgagccc tgcaagaagc ggctgactgc gcgcagcatc 3601 agcaccccca cctgcctggg gggcagccca gccgctgagc gcccggccga cctgccacca 3661 gccgccgccc tcccgcagcc cgaggtcatc ctgctagact cagacctgga tgaacccata 3721 gacttgcgct gcgtcaagac gcgcagcgag gccggggagc cgcccagctc cctccaggtg 3781 aagcccgaga caccggcgtc ggcggcggtg gcggtggcgg cggcagcggc acccaccacg 3841 acggcggaga agcctccagc cgaggcccag gacgaacctg cagagtcgct gagcgagttc 3901 aagcccttct ttgggaatat aattatcacc gacgtcaccg cgaactgcct caccgttact 3961 ttcaaggagt acgtgacggt gggatccgac tacaaagacc atgacggtga ttataaagat 4021 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc 4081 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga 4141 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat 4201 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta 4261 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga 4321 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg 4381 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag 4441 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct 4501 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt 4561 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac 4621 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac 4681 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca 4741 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4801 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4861 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4921 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4981 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 5041 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 5101 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 5161 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga 5221 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5281 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5341 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5401 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5461 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5521 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5581 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 5641 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5701 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5761 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt 5821 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5881 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5941 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6001 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6061 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6121 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6181 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6241 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6301 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6421 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6481 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6541 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6601 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6661 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6721 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6781 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6841 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6901 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6961 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7021 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7081 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7141 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7201 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7261 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7321 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7381 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7441 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7501 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7561 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7621 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7681 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7741 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7801 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7861 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7921 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7981 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8041 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8101 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8161 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8221 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8281 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8341 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8401 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8461 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8581 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8641 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8701 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8761 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8821 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8881 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8941 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 9001 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 9061 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 9121 caagctg //