LOCUS       sequence-176074-        8224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A lenti-viral vector expressing photoswitchable MEK1 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8224)
  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 8224)
  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-psMEK1
FEATURES             Location/Qualifiers
     source          1..8224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        368..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             584..764
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    811..936
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1433..1666
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1851..1895
                     /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    2193..2310
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2369..2681
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2600..2620
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2697..2713
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2698..2714
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2756..2789)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        2847..3150
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3151..3354
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3351..3375
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3480..3496
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3480..3496
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3507..3525
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3507..3524
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3548..3574
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5216..5884
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="SVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSIDLKVKE
                     GGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNATN
                     DITLDGDCYIYEIRFRGTNFPANGPVMQKRTVKWEPSTENLYVRDGVLKGDVIMALSLE
                     GGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQA
                     K"
     LTR             6230..6410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6472..7060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6552..6571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7231..8022,8023..8091))
                     /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     7854..7873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8092..8196)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
      601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
      781 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
      841 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
      901 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
      961 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1021 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1081 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1141 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1201 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1261 accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     1321 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1381 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1441 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1501 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1561 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1621 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1681 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1741 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1801 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1861 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1921 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1981 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     2041 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     2101 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     2161 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     2221 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     2281 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     2341 gtttggttag taccgggccc gctctagaga tccgacgcgc catctctagg cccgcgccgg
     2401 ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat
     2461 ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct
     2521 ttttaatact agctacattt tacatgatag gcttggattt ctataagaga tacaaatact
     2581 aaattattat tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg
     2641 tgttttgaga ctataaatat cccttggaga aaagccttgt taacgcgcgg tgaccctcga
     2701 ggtcgacggt atcgataagc tcgcttcacg agattccagc aggtcgaggg acctaataac
     2761 ttcgtatagc atacattata cgaagttata ttaagggttc caagcttaag cggccgcgtg
     2821 gataaccgta ttaccgccat gcatgtcgtt acataactta cggtaaatgg cccgcctggc
     2881 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2941 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     3001 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     3061 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     3121 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     3181 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     3241 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     3301 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     3361 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     3421 cgggaccgat ccagcctccg gactctagcc taggccgcgg gacggataac aatttcacac
     3481 aggaaacagc tatgaccatt aggcctattt aggtgacact atagaacaag tttgtacagc
     3541 caccatgtat ccctacgatg tgcccgatta cgctcccaag aagaagccga cgcccatcca
     3601 gctgaacccg acccccgatg gctcggcggt taacgggacc agctcggccg agaccaacct
     3661 ggaggccttg cagaagaagc tggaggggct ggagcttgat gagcagcagc ggaagcgcct
     3721 tgaggccttt ctgacccaga agcagaaggt gggatccggc agtagcggaa gcagtggttc
     3781 ttcaagcgtg atcaagcccg acatgaaaat caagctgagg atggagggtg ccgtgaacgg
     3841 tcacccgttt gcgatcgagg gcgtgggact gggcaagccg ttcgagggaa agcaaagcat
     3901 cgacctcaag gtgaaggagg gcggaccgct gcccttcgcc tacgacattc tgaccactgc
     3961 cttctgctac ggtaatcgcg tcttcgcaaa gtatcccgaa aacatcgtgg attactttaa
     4021 gcagagcttc ccggagggtt actcctggga gcgatccatg aactacgagg acggagggat
     4081 ctgcaacgcg acgaacgata taaccttgga cggcgactgc tacatttacg aaattcggtt
     4141 ccgcggcacc aatttcccag cgaatggccc tgtaatgcaa aaacgcacgg taaagtggga
     4201 gcccagcacc gagaacctgt acgtccgcga cggagtcctg aagggagacg tgatcatggc
     4261 gctttcattg gaaggtggcg ggcactacag atgcgatttc aagacgacct ataaggcgaa
     4321 aaaggtggtc caactgccgg actaccattt cgtcgaccac cacatcgaga tcaagagcca
     4381 cgacaaagac tatagcaacg tgaatctcca cgagcatgcc gaggcccaca gcgagctgcc
     4441 caggcaggcc aagggcctta agggttccgg taccggagaa ctgaaggatg atgactttga
     4501 gaagatcagt gagctggggg ccggcaacgg tggagtggta ttcaaggtct cccataagcc
     4561 gtctggccta gttatggcca ggaagctaat tcatctggag atcaaacccg caatccggaa
     4621 ccagatcatc cgggagctgc aggtgctgca tgagtgcaac tccccataca tcgtgggctt
     4681 ctatggggct ttctacagtg acggcgagat cagcatctgc atggaacaca tggatggtgg
     4741 gtccttggat caagttctga agaaagctgg aagaattcct gagcaaattt taggaaaagt
     4801 tagcattgct gtgataaaag gcctgacata tcttcgggag aagcacaaga ttatgcacag
     4861 agatgtcaag ccttccaata ttctagtaaa ctcccgtgga gaaatcaaac tctgtgattt
     4921 tggggtcagc gggcaactaa ttgacgacat ggccaacgac tttgtaggca caagatcata
     4981 catgtcgccc gagagactcc aggggactca ctactccgtg cagtcggaca tctggagcat
     5041 ggggctctct ctggtggaga tggcagttgg gagatacccc atccctcctc ctgatgccaa
     5101 ggaactggag ctgctgttta ccggtggatg ccaggtggag ggagatgcag ccgaaacgcc
     5161 acccaggccg aggacccctg ggaggcccct tagctcatat ggaatggaca gcggcagtgt
     5221 gattaaacca gacatgaaga tcaagctgcg tatggaaggc gctgtaaatg gacacccgtt
     5281 cgcgattgaa ggagttggcc ttgggaagcc tttcgaggga aaacagagta ttgaccttaa
     5341 agtcaaagaa ggcggacctc tgcctttcgc ctatgacatc ttgacaactg ctttctgtta
     5401 cggcaacagg gtattcgcca aatacccaga aaatatagta gactatttca agcagtcgtt
     5461 tcctgagggc tactcttggg aacgaagcat gaattacgaa gacgggggca tttgtaacgc
     5521 gacaaacgac ataaccctgg atggtgactg ttatatctat gaaattcgat ttagaggtac
     5581 gaactttcct gccaatggtc cagttatgca gaagaggact gtgaaatggg agccatccac
     5641 tgagaacttg tatgtgcgtg atggagtgct gaagggtgat gttatcatgg ctctgtcgct
     5701 tgaaggaggt ggccattacc gatgtgactt caaaactact tataaagcta agaaggttgt
     5761 ccagttgcca gactatcact ttgtggacca ccacattgag attaaaagcc acgacaaaga
     5821 ttacagtaat gttaatctgc atgagcacgc cgaagcgcat tctgagctgc cgaggcaggc
     5881 caagcgacgt acgcctccca tggcaatttt tgagttgttg gattatatag tcaatgagcc
     5941 tcctccaaaa ttgcccagtg gagtattcag tctggagttt caagattttg tgaataaatg
     6001 cttaataaag aaccctgcag agagagcaga tctaaagcag cttctggttc acgctttcat
     6061 caagagatct gatgccgagg aagtagactt cgcaggttgg ctctgctcca ccattgggct
     6121 taaccagccc agcacaccaa cccatgccgc tagcatctga acgcgtgatt ttaaaagaaa
     6181 aggggggact ggaagggcta attcactccc actgcttttt gcctgtactg ggtctctctg
     6241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc
     6421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7081 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8221 tgac
//
