LOCUS       sequence-176063-        8011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A lenti-viral vector expressing photoswitchable Raf1 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2017-02-24- 24"; volume: "355"; 
            issue: "6327"; pages: "836-842"
COMMENT     Sequence Label: pLL3.7m-psRaf1
FEATURES             Location/Qualifiers
     source          1..8011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             2069..2737
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="SVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSIDLKVKE
                     GGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNATN
                     DITLDGDCYIYEIRFRGTNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVVMALSLE
                     GGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQA
                     K"
     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(join(4180..4971,4972..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"
     promoter        7542..7854
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     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    complement(7929..7962)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
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
//
