LOCUS V023126 8944 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023126 VERSION V023126 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..3798 /label="CDC20(NM_001255)" /note="CDC20(NM_001255)" /gene="CDC20" CDS 3805..3870 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3914..4125 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4138..4406 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4433..5032 /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 5039..5627 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5701..5934 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6006..6127 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6146..6281 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6319..6335) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6343..6359 /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(6367..6397) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6412..6433 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6721..7309) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7480..8340) /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(8341..8445) /gene="bla" /label="AmpR promoter" primer_bind 8919..8935 /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 catggcacag ttcgcgttcg agagtgacct gcactcgctg 2341 cttcagctgg atgcacccat ccccaatgca ccccctgcgc gctggcagcg caaagccaag 2401 gaagccgcag gcccggcccc ctcacccatg cgggccgcca accgatccca cagcgccggc 2461 aggactccgg gccgaactcc tggcaaatcc agttccaagg ttcagaccac tcctagcaaa 2521 cctggcggtg accgctatat cccccatcgc agtgctgccc agatggaggt ggccagcttc 2581 ctcctgagca aggagaacca gcctgaaaac agccagacgc ccaccaagaa ggaacatcag 2641 aaagcctggg ctttgaacct gaacggtttt gatgtagagg aagccaagat ccttcggctc 2701 agtggaaaac cacaaaatgc gccagagggt tatcagaaca gactgaaagt actctacagc 2761 caaaaggcca ctcctggctc cagccggaag acctgccgtt acattccttc cctgccagac 2821 cgtatcctgg atgcgcctga aatccgaaat gactattacc tgaaccttgt ggattggagt 2881 tctgggaatg tactggccgt ggcactggac aacagtgtgt acctgtggag tgcaagctct 2941 ggtgacatcc tgcagctttt gcaaatggag cagcctgggg aatatatatc ctctgtggcc 3001 tggatcaaag agggcaacta cttggctgtg ggcaccagca gtgctgaggt gcagctatgg 3061 gatgtgcagc agcagaaacg gcttcgaaat atgaccagtc actctgcccg agtgggctcc 3121 ctaagctgga acagctatat cctgtccagt ggttcacgtt ctggccacat ccaccaccat 3181 gatgttcggg tagcagaaca ccatgtggcc acactgagtg gccacagcca ggaagtgtgt 3241 gggctgcgct gggccccaga tggacgacat ttggccagtg gtggtaatga taacttggtc 3301 aatgtgtggc ctagtgctcc tggagagggt ggctgggttc ctctgcagac attcacccag 3361 catcaagggg ctgtcaaggc cgtagcatgg tgtccctggc agtccaatgt cctggcaaca 3421 ggagggggca ccagtgatcg acacattcgc atctggaatg tgtgctctgg ggcctgtctg 3481 agtgccgtgg atgcccattc ccaggtgtgc tccatcctct ggtctcccca ttacaaggag 3541 ctcatctcag gccatggctt tgcacagaac cagctagtta tttggaagta cccaaccatg 3601 gccaaggtgg ctgaactcaa aggtcacaca tcccgggtcc tgagtctgac catgagccca 3661 gatggggcca cagtggcatc cgcagcagca gatgagaccc tgaggctatg gcgctgtttt 3721 gagttggacc ctgcgcggcg gcgggagcgg gagaaggcca gtgcagccaa aagcagcctc 3781 atccaccaag gcatccgcgg atccgactac aaagaccatg acggtgatta taaagatcat 3841 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 3901 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 3961 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4021 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4081 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4141 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4201 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4261 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4321 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4381 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4441 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4501 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4561 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4621 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4681 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4741 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4801 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4861 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4921 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4981 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5041 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5101 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5161 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5221 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5281 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5341 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5401 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5461 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5521 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5581 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5641 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5701 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5761 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5821 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5881 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5941 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6001 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6061 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6121 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6181 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6241 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6301 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6361 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6421 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6481 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6541 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6601 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6661 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6721 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6781 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6841 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6901 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6961 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7021 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7081 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7141 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7201 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7261 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7321 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7381 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7441 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7501 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7561 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7621 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7681 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7741 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7801 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7861 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7921 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7981 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8041 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8101 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8161 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8221 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8281 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8341 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8401 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8461 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8521 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8581 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8641 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8701 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8761 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8821 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8881 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8941 gctg //