LOCUS V031546 9704 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031546 VERSION V031546 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 2290..3846 /label="STK3(NM_001256312)" /note="STK3(NM_001256312)" /gene="STK3" CDS 3883..4599 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 4606..4623 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 4674..4885 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4898..5166 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5193..5792 /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 5799..6387 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6461..6694 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6766..6887 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6906..7041 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7079..7095) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7103..7119 /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(7127..7157) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7172..7193 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7481..8069) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8240..9100) /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(9101..9205) /gene="bla" /label="AmpR promoter" primer_bind 9679..9695 /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 agagccacca tgttacaact catggacagt ggcattacca tctgccttag gaacggagca 2341 gcctctgtct tcaagaaaaa agaatggagc acacaaggag aagaaaacaa gcaagacagt 2401 aaactaaaaa agctgagtga agacagtttg actaagcagc ctgaagaagt ttttgatgta 2461 ttagagaagc ttggagaagg gtcttatgga agtgtattta aagcaataca caaggaatcc 2521 ggtcaagttg tcgcaattaa acaagtacct gttgaatcag atcttcagga aataatcaaa 2581 gaaatttcca taatgcagca atgtgacagc ccatatgttg taaagtacta tggcagttat 2641 tttaagaata cagacctctg gattgttatg gagtactgtg gcgctggctc tgtctcagac 2701 ataattagat tacgaaacaa gacattaata gaagatgaaa ttgcaaccat tcttaaatct 2761 acattgaaag gactagaata tttgcacttt atgagaaaaa tacacagaga tataaaagct 2821 ggaaatattc tcctcaatac agaaggacat gcaaaattgg cagattttgg agtggctggt 2881 cagttaacag atacaatggc aaaacgcaat actgtaatag gaactccatt ttggatggct 2941 cctgaggtga ttcaagaaat aggctataac tgtgtggccg acatctggtc ccttggcatt 3001 acttctatag aaatggctga aggaaaacct ccttatgctg atatacatcc aatgagggct 3061 atttttatga ttcccacaaa tccaccacca acattcagaa agccagaact ttggtccgat 3121 gatttcaccg attttgttaa aaagtgtttg gtgaagaatc ctgagcagag agctactgca 3181 acacaacttt tacagcatcc ttttatcaag aatgccaaac ctgtatcaat attaagagac 3241 ctgatcacag aagctatgga gatcaaagct aaaagacatg aggaacagca acgagaattg 3301 gaagaggaag aagaaaattc ggatgaagat gagctggatt cccacaccat ggtgaagact 3361 agtgtggaga gtgtgggcac catgcgggcc acaagcacga tgagtgaagg ggcccagacc 3421 atgattgaac ataatagcac gatgttggaa tccgacttgg ggaccatggt gataaacagt 3481 gaggatgagg aagaagaaga tggaactatg aaaagaaatg caacctcacc acaagtacaa 3541 agaccatctt tcatggacta ctttgataag caagacttca agaataagag tcacgaaaac 3601 tgtaatcaga acatgcatga acccttccct atgtccaaaa acgtttttcc tgataactgg 3661 aaagttcctc aagatggaga ctttgacttt ttgaaaaatc taagtttaga agaactacag 3721 atgcggttaa aagcactgga ccccatgatg gaacgggaga tagaagaact tcgtcagaga 3781 tacactgcga aaagacagcc cattctggat gcgatggatg caaagaaaag aaggcagcaa 3841 aactttgaat tcggaggtgg cggaagcggc ggtggaggta gtatggtgag caagggcgag 3901 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac 3961 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag 4021 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc 4081 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag 4141 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac 4201 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg 4261 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac 4321 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc 4381 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac 4441 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc 4501 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc 4561 gccgccggga tcactctcgg catggacgag ctgtacaagc tcgagcacca ccaccaccac 4621 cactgaggat ccgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga 4681 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc 4741 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt 4801 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg 4861 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg 4921 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc 4981 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag 5041 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct 5101 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta 5161 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc 5221 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact 5281 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc 5341 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 5401 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 5461 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 5521 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 5581 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 5641 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 5701 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 5761 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa 5821 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 5881 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 5941 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 6001 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 6061 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 6121 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 6181 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc 6241 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 6301 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 6361 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 6421 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 6481 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 6541 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6601 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6661 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6721 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6901 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6961 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 7021 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 7081 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7141 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7201 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7921 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 8041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 8101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 9001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 9061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 9241 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 9301 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 9361 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 9421 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 9481 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 9541 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 9601 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 9661 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //