LOCUS       sequence-176060-        8245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A lenti-viral vector expressing photoswitchable MEK2 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8245)
  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 8245)
  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-psMEK2
FEATURES             Location/Qualifiers
     source          1..8245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        34..337
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        338..541
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..562
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     667..683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     667..683
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        694..712
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     694..711
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             735..761
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2427..3095
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="SVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSIDLKVKE
                     GGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNATN
                     DITLDGDCYIYEIRFRGTNFPANGPVMQKRTVKWEPSTENLYVRDGVLKGDVIMALSLE
                     GGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQA
                     K"
     LTR             3438..3618
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3680..4268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3760..3779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4439..5230,5231..5299))
                     /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     5062..5081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5300..5404)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5495..5798
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5800..5998
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6016..6196
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6243..6368
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6865..7098
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7283..7327
                     /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    7625..7742
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7801..8113
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     8032..8052
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     8129..8145
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8130..8146
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8188..8221)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ccgcgtggat aaccgtatta ccgccatgca tgtcgttaca taacttacgg taaatggccc
       61 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      121 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      181 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      241 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      301 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      361 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      421 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      481 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      541 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      601 aagacaccgg gaccgatcca gcctccggac tctagcctag gccgcgggac ggataacaat
      661 ttcacacagg aaacagctat gaccattagg cctatttagg tgacactata gaacaagttt
      721 gtacagccac catgtatccc tacgatgtgc ccgattacgc tcttgccagg aggaagcctg
      781 tgctgcccgc tcttacaatt aatcctacca ttgccgaggg accaagccct accagcgaag
      841 gcgcatccga agctaatctg gtggacctgc agaagaaact tgaagaactt gagcttgatg
      901 agcagcagaa aaagaggctg gaggcgttcc tcacacagaa ggcaaaagtg ggatccggca
      961 gtagcggaag cagtggttct tcaagcgtga tcaagcccga catgaaaatc aagctgagga
     1021 tggagggtgc cgtgaacggt cacccgtttg cgatcgaggg cgtgggactg ggcaagccgt
     1081 tcgagggaaa gcaaagcatc gacctcaagg tgaaggaggg cggaccgctg cccttcgcct
     1141 acgacattct gaccactgcc ttctgctacg gtaatcgcgt cttcgcaaag tatcccgaaa
     1201 acatcgtgga ttactttaag cagagcttcc cggagggtta ctcctgggag cgatccatga
     1261 actacgagga cggagggatc tgcaacgcga cgaacgatat aaccttggac ggcgactgct
     1321 acatttacga aattcggttc cgcggcacca atttcccagc gaatggccct gtaatgcaaa
     1381 aacgcacggt aaagtgggag cccagcaccg agaacctgta cgtccgcgac ggagtcctga
     1441 agggagacgt gatcatggcg ctttcattgg aaggtggcgg gcactacaga tgcgatttca
     1501 agacgaccta taaggcgaaa aaggtggtcc aactgccgga ctaccatttc gtcgaccacc
     1561 acatcgagat caagagccac gacaaagact atagcaacgt gaatctccac gagcatgccg
     1621 aggcccacag cgagctgccc aggcaggcca agggccttaa gggatccggt accggggagc
     1681 tgaaagatga tgattttgag agaatatcag agcttggcgc aggtaatggc ggcgtggtaa
     1741 ctaaggtgca gcaccgccca tcaggactca ttatggctag aaagctcatt caccttgaga
     1801 taaagcctgc tatacgcaac cagataattc gggagctgca ggtcctgcac gagtgtaatt
     1861 caccctacat tgttggcttc tacggggcat tttactcaga cggagagatt tctatatgca
     1921 tggagcacat ggacggagga agcctggacc aggtcctgaa ggaagcaaag aggattcccg
     1981 aggagatcct gggcaaggtg agcattgctg tgctgcgcgg gttggcatac ctgcgggaaa
     2041 agcaccagat catgcatcgg gacgtcaaac ctagcaacat cttggtgaat agtcggggcg
     2101 aaattaaact ttgtgatttc ggtgtgtccg gccagttgat tgacgatatg gctaatgatt
     2161 tcgtcggcac ccgctcctac atggctccgg aaaggctcca gggaacgcac tacagcgtgc
     2221 agtcagacat ctggtccatg gggctgagcc tcgtggaatt ggcggtaggc agatacccta
     2281 taccaccacc tgatgcaaaa gagctggagg ccatcttcgg cagacccgtg gttgacgggg
     2341 aggagggtga accacactcc attagcccaa gacctcgacc accagggagg cctgtaagtg
     2401 ggcacggaat ggacagcacc ggtggcagtg tgattaaacc agacatgaag atcaagctgc
     2461 gtatggaagg cgctgtaaat ggacacccgt tcgcgattga aggagttggc cttgggaagc
     2521 ctttcgaggg aaaacagagt attgacctta aagtcaaaga aggcggacct ctgcctttcg
     2581 cctatgacat cttgacaact gctttctgtt acggcaacag ggtattcgcc aaatacccag
     2641 aaaatatagt agactatttc aagcagtcgt ttcctgaggg ctactcttgg gaacgaagca
     2701 tgaattacga agacgggggc atttgtaacg cgacaaacga cataaccctg gatggtgact
     2761 gttatatcta tgaaattcga tttagaggta cgaactttcc tgccaatggt ccagttatgc
     2821 agaagaggac tgtgaaatgg gagccatcca ctgagaactt gtatgtgcgt gatggagtgc
     2881 tgaagggtga tgttatcatg gctctgtcgc ttgaaggagg tggccattac cgatgtgact
     2941 tcaaaactac ttataaagct aagaaggttg tccagttgcc agactatcac tttgtggacc
     3001 accacattga gattaaaagc cacgacaaag attacagtaa tgttaatctg catgagcacg
     3061 ccgaagcgca ttctgagctg ccgaggcagg ccaagcgtac gaggcccgcc atggctatct
     3121 ttgaactgct ggattacatc gtgaacgagc ctccacccaa gctgcccaac ggtgttttca
     3181 cacccgactt ccaggaattc gtcaataaat gcctgatcaa aaatccagca gagagagctg
     3241 acctcaagat gctgacaaac cacacgttta ttaaaaggtc cgaagttgag gaagtcgatt
     3301 tcgctggctg gctgtgcaag acgctcaggc tgaatcagcc cggaactccc actagaacag
     3361 cagtttgaac gcgtgatttt aaaagaaaag gggggactgg aagggctaat tcactcccac
     3421 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     3481 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3541 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3601 tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4081 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4561 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5401 cgcgcacatt tccccgaaaa gtgccacctg acactagtta ttaatagtaa tcaattacgg
     5461 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     5521 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     5581 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     5641 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     5701 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     5761 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     5821 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     5881 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5941 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg
     6001 cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6061 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6121 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6181 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     6241 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     6301 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     6361 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     6421 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     6481 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     6541 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     6601 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     6661 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
     6721 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     6781 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     6841 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     6901 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     6961 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     7021 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     7081 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     7141 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     7201 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     7261 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     7321 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     7381 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     7441 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     7501 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     7561 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     7621 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     7681 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     7741 ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga
     7801 gatccgacgc gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc
     7861 tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct
     7921 taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat
     7981 aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa
     8041 aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga
     8101 gaaaagcctt gttaacgcgc ggtgaccctc gaggtcgacg gtatcgataa gctcgcttca
     8161 cgagattcca gcaggtcgag ggacctaata acttcgtata gcatacatta tacgaagtta
     8221 tattaagggt tccaagctta agcgg
//
