LOCUS Exported 8197 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A lenti-viral vector expressing photoswitchable MEK1 with minimum basal activity in mammalian cells. ACCESSION . VERSION . KEYWORDS pLL3.7m-psMEK1tight SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8197) AUTHORS Zhou XX, Fan LZ, Li P, Shen K, Lin MZ TITLE Optical control of cell signaling by single-chain photoswitchable kinases. JOURNAL Science. 2017 Feb 24;355(6327):836-842. doi: 10.1126/science.aah3605. PUBMED 28232577 REFERENCE 2 (bases 1 to 8197) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8197 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 30..333 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 335..533 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 551..731 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 778..903 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1400..1633 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1818..1862 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2160..2277 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" primer_bind 2567..2587 /label=mU6-F /note="Mouse U6 promoter, forward primer" primer_bind 2664..2680 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2665..2681 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 2723..2756 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." enhancer 2814..3117 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3118..3321 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3318..3342 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 3447..3463 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3447..3463 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 3474..3492 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 3474..3491 /label=SP6 /note="SP6 promoter, forward primer" CDS 3515..3541 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" LTR 6170..6350 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(6412..7000) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6492..6511) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7171..8031) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7794..7813 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8032..8136) /gene="bla" /label=AmpR promoter ORIGIN 1 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 61 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 121 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 181 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 241 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 301 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 361 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 421 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 481 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt 541 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 601 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 661 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 721 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc 781 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg 841 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc 901 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg 961 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc 1021 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta 1081 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc 1141 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata 1201 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca 1261 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag 1321 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag 1381 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca 1441 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag 1501 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca 1561 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg 1621 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc 1681 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga 1741 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat 1801 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt 1861 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag 1921 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta 1981 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca 2041 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag 2101 tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat 2161 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata 2221 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg 2281 gtttattaca gggacagcag agatccagtt tggttagtac cgggcccgct ctagagatcc 2341 gacgcgccat ctctaggccc gcgccggccc cctcgcacag acttgtggga gaagctcggc 2401 tactcccctg ccccggttaa tttgcatata atatttccta gtaactatag aggcttaatg 2461 tgcgataaaa gacagataat ctgttctttt taatactagc tacattttac atgataggct 2521 tggatttcta taagagatac aaatactaaa ttattatttt aaaaaacagc acaaaaggaa 2581 actcacccta actgtaaagt aattgtgtgt tttgagacta taaatatccc ttggagaaaa 2641 gccttgttaa cgcgcggtga ccctcgaggt cgacggtatc gataagctcg cttcacgaga 2701 ttccagcagg tcgagggacc taataacttc gtatagcata cattatacga agttatatta 2761 agggttccaa gcttaagcgg ccgcgtggat aaccgtatta ccgccatgca tgtcgttaca 2821 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 2881 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 2941 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 3001 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 3061 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 3121 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 3181 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 3241 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 3301 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca 3361 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac tctagcctag 3421 gccgcgggac ggataacaat ttcacacagg aaacagctat gaccattagg cctatttagg 3481 tgacactata gaacaagttt gtacagccac catgtatccc tacgatgtgc ccgattacgc 3541 tcccaagaag aagccgacgc ccatccagct gaacccgacc cccgatggct cggcggttaa 3601 cgggaccagc tcggccgaga ccaacctgga ggccttgcag aagaagctgg aggggctgga 3661 gcttgatgag cagcagcgga agcgccttga ggcctttctg acccagaagc agaaggtggg 3721 atccggcagt agcggaagca gtggttcttc aagcgtgatc aagcccgaca tgaaaatcaa 3781 gctgaggatg gagggtgccg tgaacggtca cccgtttgcg atcgagggcg tgggactggg 3841 caagccgttc gagggaaagc aaagcatcga cctcaaggtg aaggagggcg gaccgctgcc 3901 cttcgcctac gacattctga ccactgcctt ctgctacggt aatcgcgtct tcgcaaagta 3961 tcccgaaaac atcgtggatt actttaagca gagcttcccg gagggttact cctgggagcg 4021 atccatgaac tacgaggacg gagggatctg caacgcgacg aacgatataa ccttggacgg 4081 cgactgctac atttacgaaa ttcggttccg cggcaccaat ttcccagcga atggccctgt 4141 aatgcaaaaa cgcacggtaa agtgggagcc cagcaccgag aacctgtacg tccgcgacgg 4201 agtcctgaag ggagacgtga tcatggcgct ttcattggaa ggtggcgggc actacagatg 4261 cgatttcaag acgacctata aggcgaaaaa ggtggtccaa ctgccggact accatttcgt 4321 cgaccaccac atcgagatca agagccacga caaagactat agcaacgtga atctccacga 4381 gcatgccgag gcccacagcg agctgcccag gcagggagaa ctgaaggatg atgactttga 4441 gaagatcagt gagctggggg ccggcaacgg tggagtggta ttcaaggtct cccataagcc 4501 gtctggccta gttatggcca ggaagctaat tcatctggag atcaaacccg caatccggaa 4561 ccagatcatc cgggagctgc aggtgctgca tgagtgcaac tccccataca tcgtgggctt 4621 ctatggggct ttctacagtg acggcgagat cagcatctgc atggaacaca tggatggtgg 4681 gtccttggat caagttctga agaaagctgg aagaattcct gagcaaattt taggaaaagt 4741 tagcattgct gtgataaaag gcctgacata tcttcgggag aagcacaaga ttatgcacag 4801 agatgtcaag ccttccaata ttctagtaaa ctcccgtgga gaaatcaaac tctgtgattt 4861 tggggtcagc gggcaactaa ttgacgacat ggccaacgac tttgtaggca caagatcata 4921 catgtcgccc gagagactcc aggggactca ctactccgtg cagtcggaca tctggagcat 4981 ggggctctct ctggtggaga tggcagttgg gagatacccc atccctcctc ctgatgccaa 5041 ggaactggag ctgctgttta ccggtggatg ccaggtggag ggagatgcag ccgaaacgcc 5101 acccaggccg aggacccctg ggaggcccct tagctcatat ggaatggaca gcggcagtgt 5161 gattaaacca gacatgaaga tcaagctgcg tatggaaggc gctgtaaatg gacacccgtt 5221 cgcgattgaa ggagttggcc ttgggaagcc tttcgaggga aaacagagta ttgaccttaa 5281 agtcaaagaa ggcggacctc tgcctttcgc ctatgacatc ttgacaactg ctttctgtta 5341 cggcaacagg gtattcgcca aatacccaga aaatatagta gactatttca agcagtcgtt 5401 tcctgagggc tactcttggg aacgaagcat gaattacgaa gacgggggca tttgtaacgc 5461 gacaaacgac ataaccctgg atggtgactg ttatatctat gaaattcgat ttagaggtac 5521 gaactttcct gccaatggtc cagttatgca gaagaggact gtgaaatggg agccatccac 5581 tgagaacttg tatgtgcgtg atggagtgct gaagggtgat gttatcatgg ctctgtcgct 5641 tgaaggaggt ggccattacc gatgtgactt caaaactact tataaagcta agaaggttgt 5701 ccagttgcca gactatcact ttgtggacca ccacattgag attaaaagcc acgacaaaga 5761 ttacagtaat gttaatctgc atgagcacgc cgaagcgcat tctgagctgc cgaggcaggc 5821 caagcgacgt acgcctccca tggcaatttt tgagttgttg gattatatag tcaatgagcc 5881 tcctccaaaa ttgcccagtg gagtattcag tctggagttt caagattttg tgaataaatg 5941 cttaataaag aaccctgcag agagagcaga tctaaagcag cttctggttc acgctttcat 6001 caagagatct gatgccgagg aagtagactt cgcaggttgg ctctgctcca ccattgggct 6061 taaccagccc agcacaccaa cccatgccgc tagcatctga acgcgtgatt ttaaaagaaa 6121 aggggggact ggaagggcta attcactccc actgcttttt gcctgtactg ggtctctctg 6181 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6241 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6301 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc 6361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 6421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 6481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6601 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6841 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 6961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 7021 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 7081 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 7141 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 7201 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 7261 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 7321 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 7381 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 7441 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 7501 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7561 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7621 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7681 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7741 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7801 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7861 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7921 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 7981 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 8041 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 8101 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 8161 tgacactagt tattaatagt aatcaattac ggggtca //