LOCUS       sequence-176059-        8197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A lenti-viral vector expressing photoswitchable MEK1 with minimum 
            basal activity in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 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-psMEK1tight
FEATURES             Location/Qualifiers
     source          1..8197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        2336..2648
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     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    complement(2723..2756)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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"
     CDS             5156..5824
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="SVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSIDLKVKE
                     GGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNATN
                     DITLDGDCYIYEIRFRGTNFPANGPVMQKRTVKWEPSTENLYVRDGVLKGDVIMALSLE
                     GGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQA
                     K"
     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(join(7171..7962,7963..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
//
