LOCUS Exported 8011 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A lenti-viral vector expressing photoswitchable Raf1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pLL3.7m-psRaf1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8011) 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 8011) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8011 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 9..312 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 313..516 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 513..537 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 642..658 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 642..658 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 669..687 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 669..686 /label=SP6 /note="SP6 promoter, forward primer" CDS 710..736 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" LTR 3179..3359 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(3421..4009) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3501..3520) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4180..5040) /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 4803..4822 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5041..5145) /gene="bla" /label=AmpR promoter enhancer 5236..5539 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5541..5739 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 5757..5937 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5984..6109 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6606..6839 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7024..7068 /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 7366..7483 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" primer_bind 7773..7793 /label=mU6-F /note="Mouse U6 promoter, forward primer" primer_bind 7870..7886 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 7871..7887 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 7929..7962 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." ORIGIN 1 atgcatgtcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 481 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc 541 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc 601 cggactctag cctaggccgc gggacggata acaatttcac acaggaaaca gctatgacca 661 ttaggcctat ttaggtgaca ctatagaaca agtttgtaca gccaccatgt atccctacga 721 tgtgcccgat tacgctagcg tgatcaagcc cgacatgaaa atcaagctga ggatggaggg 781 tgccgtgaac ggtcacccgt ttgcgatcga gggcgtggga ctgggcaagc cgttcgaggg 841 aaagcaaagc atcgacctca aggtgaagga gggcggaccg ctgcccttcg cctacgacat 901 tctgaccact gccttctgct acggtaatcg cgtcttcgca aagtatcccg aaaacatcgt 961 ggattacttt aagcagagct tcccggaggg ttactcctgg gagcgatcca tgaactacga 1021 ggacggaggg atctgcaacg cgacgaacga tataaccttg gacggcgact gctacattta 1081 cgaaattcgg ttccgcggca ccaatttccc agcgaatggc cctgtaatgc aaaaacgcac 1141 ggtaaagtgg gagcccagca ccgagaacct gtacgtccgc gacggagtcc tgaagggaga 1201 cgtggtcatg gcgctttcat tggaaggtgg cgggcactac agatgcgatt tcaagacgac 1261 ctataaggcg aaaaaggtgg tccaactgcc ggactaccat ttcgtcgacc accacatcga 1321 gatcaagagc cacgacaaag actatagcaa cgtgaatctc cacgagcatg ccgaggccca 1381 cagcgagctg cccaggcagg ccaagggcct taagggatcc tattattggg aaatagaagc 1441 cagtgaagtg atgctgtcca ctcggattgg gtcaggctct tttggaactg tttataaggg 1501 taaatggcac ggagatgttg cagtaaagat cctaaaggtt gtcgacccaa ccccagagca 1561 attccaggcc ttcaggaatg aggtggctgt tctgcgcaaa acacggcatg tgaacattct 1621 gcttttcatg gggtacatga caaaggacaa cctggcaatt gtgacccagt ggtgcgaggg 1681 cagcagcctc tacaaacacc tgcatgtcca ggagaccaag tttcagatgt tccagctaat 1741 tgacattgcc cggcagacgg ctcagggaat ggactatttg catgcaaaga acatcatcca 1801 tagagacatg aaatccaaca atatatttct ccatgaaggc ttaacagtga aaattggaga 1861 ttttggtttg gcaacagtaa agtcacgctg gagtggttct cagcaggttg aacaacctac 1921 tggctctgtc ctctggatgg ccccagaggt gatccgaatg caggataaca acccattcag 1981 tttccagtcg gatgtctact cctatggcat cgtattgtat gaactgatga cgggggagct 2041 tccttattct cacatcaaca ccggtggcag tgtgattaaa ccagacatga agatcaagct 2101 gcgtatggaa ggcgctgtaa atggacaccc gttcgcgatt gaaggagttg gccttgggaa 2161 gcctttcgag ggaaaacaga gtattgacct taaagtcaaa gaaggcggac ctctgccttt 2221 cgcctatgac atcttgacaa ctgctttctg ttacggcaac agggtattcg ccaaataccc 2281 agaaaatata gtagactatt tcaagcagtc gtttcctgag ggctactctt gggaacgaag 2341 catgaattac gaagacgggg gcatttgtaa cgcgacaaac gacataaccc tggatggtga 2401 ctgttatatc tatgaaattc gatttagagg tacgaacttt cctgccaatg gtccagttat 2461 gcagaagagg actgtgaaat gggagccatc cactgagaaa ttgtatgtgc gtgatggagt 2521 gctgaagggt gatgttgtca tggctctgtc gcttgaagga ggtggccatt accgatgtga 2581 cttcaaaact acttataaag ctaagaaggt tgtccagttg ccagactatc actttgtgga 2641 ccaccacatt gagattaaaa gccacgacaa agattacagt aatgttaatc tgcatgagca 2701 cgccgaagcg cattctgagc tgccgaggca ggccaagcgt acgaaccgag atcagatcat 2761 cttcatggtg ggccgaggat atgcctcccc agatcttagt aagctatata agaactgccc 2821 caaagcaatg aagaggctgg tagctgactg tgtgaagaaa gtaaaggagg agaggcctct 2881 ttttccccag atcctgtctt ccattgagct gctccaacac tctctaccga agatcaaccg 2941 gagcgcttcc gagccatcct tgcatcgggc agcccacact gaggatatca atgcttgcac 3001 gctgaccacg tccccgaggc tgcctgtctt cgctagcgaa ttcggcaaga tgagcaagga 3061 cggcaagaag aaaaagaaga agtccaagac taagtgcgtg atcatgtgaa cgcgtgattt 3121 taaaagaaaa ggggggactg gaagggctaa ttcactccca ctgctttttg cctgtactgg 3181 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 3241 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 3301 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 3361 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 3421 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 3481 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 3541 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 3601 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 3661 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 3721 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 3781 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 3841 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 3901 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 3961 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 4021 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 4081 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 4141 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 4201 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 4261 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 4321 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 4381 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 4441 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 4501 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 4561 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 4621 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 4681 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 4741 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 4801 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 4861 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 4921 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 4981 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 5041 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 5101 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 5161 agtgccacct gacactagtt attaatagta atcaattacg gggtcattag ttcatagccc 5221 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 5281 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 5341 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 5401 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 5461 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 5521 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 5581 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 5641 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 5701 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt 5761 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5821 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5881 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg 5941 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac 6001 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa 6061 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg 6121 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata 6181 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg 6241 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca 6301 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca 6361 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa 6421 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga 6481 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 6541 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 6601 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 6661 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 6721 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 6781 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 6841 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 6901 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 6961 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 7021 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 7081 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 7141 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 7201 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata 7261 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca 7321 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg 7381 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca 7441 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga 7501 cagcagagat ccagtttggt tagtaccggg cccgctctag agatccgacg cgccatctct 7561 aggcccgcgc cggccccctc gcacagactt gtgggagaag ctcggctact cccctgcccc 7621 ggttaatttg catataatat ttcctagtaa ctatagaggc ttaatgtgcg ataaaagaca 7681 gataatctgt tctttttaat actagctaca ttttacatga taggcttgga tttctataag 7741 agatacaaat actaaattat tattttaaaa aacagcacaa aaggaaactc accctaactg 7801 taaagtaatt gtgtgttttg agactataaa tatcccttgg agaaaagcct tgttaacgcg 7861 cggtgaccct cgaggtcgac ggtatcgata agctcgcttc acgagattcc agcaggtcga 7921 gggacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg ttccaagctt 7981 aagcggccgc gtggataacc gtattaccgc c //