LOCUS       sequence-401663-        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes engineered opto-ZF9 with AsLOV2(408-543) inserted in C2 
            position in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  AUTHORS   Zhu L, McNamara HM, Toettcher JE
  TITLE     Light-switchable transcription factors obtained by direct screening 
            in mammalian cells.
  JOURNAL   Nat Commun. 2023 Jun 2;14(1):3185. doi: 10.1038/s41467-023-38993-6.
  PUBMED    37268649
REFERENCE   2  (bases 1 to 7897)
  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"; journalName: "Nat 
            Commun."; date: "2023-06-2"; volume: "14(1):3185. doi"; pages: " 
            10.1038/s41467-023-38993-6"
COMMENT     Sequence Label: pEF-1-alpha_ZF9LOV[C2]-VP64 (pLZA126)
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(74..94)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        256..1562
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          583..1530
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     CDS             1585..1608
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1609..1629
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2329..2478
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     polyA_signal    2510..2631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2521..2540)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2575..2594
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2819..3407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3308..3327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3487..3790
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3905..5210
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    4247..5178
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             5223..5933
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWQASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5368..5386)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5636..5655
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5940..6384
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        6417..6463
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6455..6473
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6483..7277
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6483..7277
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6537..6556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7147..7166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7324..7896
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 cctgcagggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
       61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      241 ccactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag
      301 cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg
      361 cgaggagcat gcgctcaggg ctgagcgcgg ggagagcaga gcacacaagc tcatagaccc
      421 tggtcgtggg ggggaggacc ggggagctgg cgcggggcaa actgggaaag cggtgtcgtg
      481 tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc ggcagtcgcc
      541 ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagggg cgggtgtggc
      601 ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg ccgggccccg ctgtcgtcgg
      661 cggggattag ctgcgagcat tcccgcttcg agttgcgggc ggcgcgggag gcagagtgcg
      721 aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc
      781 cgcgccgccg ccgcgtgcta ctccggccgc actctggtct tttttttttt tgttgttgtt
      841 gccctgctgc cttcgattgc cgttcagcaa taggggctaa caaagggagg gtgcggggct
      901 tgctcgcccg gagcccggag aggtcatggt tggggaggaa tggagggaca ggagtggcgg
      961 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgg
     1021 ggtttttccc gaagcaacca ggctggggtt agcgtgccga ggccatgtgg ccccagcacc
     1081 cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc cctaactagg gtgaggccat
     1141 cccgtccggc accagttgcg tgcgtggaaa gatggccgct cccgggccct gttgcaagga
     1201 gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg
     1261 aagagggcct ggtccctcac cggctgctgc ttcctgtgac cccgtggtcc tatcggccgc
     1321 aatagtcacc tcgggctttt gagcacggct agtcgcggcg gggggagggg atgtaatggc
     1381 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     1441 catttttgga atttgtcccc ttgagttttg agcggagcta attctcgggc ttcttagcgg
     1501 ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa aaccaccgct aattcaaagc
     1561 aaccggtgat atcaaagatc tatggactac aaggacgacg atgacaagcc caagaaaaag
     1621 cgcaaggtct ctagacccgg ggagcgcccc ttccagtgtc gcatttgcat ggcatctttg
     1681 gaacgtatcg aaaagaattt tgtcatcacg gatccgcgtc ttcccgacaa tccgattatc
     1741 ttcgcgtcag actctttctt acaactgact gagtatagta gagaggagat attggggcgt
     1801 aactgtagat ttcttcaggg gccagaaact gatcgggcta ccgttcgcaa gatacgtgac
     1861 gcaatagaca accagaccga ggtgacggtg cagctgatta actacacaaa gtctgggaag
     1921 aagttctgga acctgtttca tttacaacct atgagagatc aaaaaggtga cgttcaatat
     1981 ttcatcgggg ttcagttaga tgggactgag cacgtgagag atgcagcaga aagagagggt
     2041 gtaatgctta ttaaaaaaac agccgagaat atcgacgaag ccgctgcgtc acggaacttt
     2101 tcggataaaa ctaaattgag agttcatacc cgtactcata ccggtgaaaa accgtttcag
     2161 tgtcggatct gtatgcgaaa tttctccgtt agacataatt tgactagaca tctacgtacg
     2221 cacaccggcg agaagccatt ccaatgccga atatgcatgc gcaacttcag tcaatctact
     2281 tctttgcaaa gacacctaaa aacccacctg agaggatccg ggcgcgccga cgcgctggac
     2341 gatttcgatc tcgacatgct gggttctgat gccctcgatg actttgacct ggatatgttg
     2401 ggaagcgacg cattggatga ctttgatctg gacatgctcg gctccgatgc tctggacgat
     2461 ttcgatctcg atatgttaat taattgttaa gctagctggc cagacatgat aagatacatt
     2521 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2581 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2641 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2701 taaaacctct acaaatgtgg tatggaaatg ttaattaact agccatgacc aaaatccctt
     2761 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2821 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2881 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2941 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3001 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3061 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3121 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3181 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3241 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3301 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3361 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3421 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc ttaattaacc
     3481 tgcaggcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3541 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3601 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3661 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3721 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3781 tattaccatg atgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     3841 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt gactagtgga
     3901 gccgagagta attcatacaa aaggagggat cgccttcgca aggggagagc ccagggaccg
     3961 tccctaaatt ctcacagacc caaatccctg tagccgcccc acgacagcgc gaggagcatg
     4021 cgcccagggc tgagcgcggg tagatcagag cacacaagct cacagtcccc ggcggtgggg
     4081 ggaggggcgc gctgagcggg ggccagggag ctggcgcggg gcaaactggg aaagtggtgt
     4141 cgtgtgctgg ctccgccctc ttcccgaggg tgggggagaa cggtatataa gtgcggtagt
     4201 cgccttggac gttctttttc gcaacgggtt tgccgtcaga acgcaggtga gtggcgggtg
     4261 tggcttccgc gggccccgga gctggagccc tgctctgagc gggccgggct gatatgcgag
     4321 tgtcgtccgc agggtttagc tgtgagcatt cccacttcga gtggcgggcg gtgcgggggt
     4381 gagagtgcga ggcctagcgg caaccccgta gcctcgcctc gtgtccggct tgaggcctag
     4441 cgtggtgtcc gccgccgcgt gccactccgg ccgcactatg cgttttttgt ccttgctgcc
     4501 ctcgattgcc ttccagcagc atgggctaac aaagggaggg tgtggggctc actcttaagg
     4561 agcccatgaa gcttacgttg gataggaatg gaagggcagg aggggcgact ggggcccgcc
     4621 cgccttcgga gcacatgtcc gacgccacct ggatggggcg aggcctgtgg ctttccgaag
     4681 caatcgggcg tgagtttagc ctacctgggc catgtggccc tagcactggg cacggtctgg
     4741 cctggcggtg ccgcgttccc ttgcctccca acaagggtga ggccgtcccg cccggcacca
     4801 gttgcttgcg cggaaagatg gccgctcccg gggccctgtt gcaaggagct caaaatggag
     4861 gacgcggcag cccggtggag cgggcgggtg agtcacccac acaaaggaag agggccttgc
     4921 ccctcgccgg ccgctgcttc ctgtgacccc gtggtctatc ggccgcatag tcacctcggg
     4981 cttctcttga gcaccgctcg tcgcggcggg gggaggggat ctaatggcgt tggagtttgt
     5041 tcacatttgg tgggtggaga ctagtcaggc cagcctggcg ctggaagtca ttcttggaat
     5101 ttgccccttt gagtttggag cgaggctaat tctcaagcct cttagcggtt caaaggtatt
     5161 ttctaaaccc gtttccaggt gttgtgaaag ccaccgctaa ttcaaagcaa tccggagtat
     5221 acatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     5281 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     5341 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     5401 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     5461 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     5521 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     5581 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     5641 taatgcagaa gaagaccatg ggctggcagg cctcctccga gcggatgtac cccgaggacg
     5701 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     5761 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     5821 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     5881 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taacctagga
     5941 gcaggtttcc ccaatgacac aaaacgtgca acttgaaact ccgcctggtc tttccaggtc
     6001 tagaggggta acactttgta ctgcgtttgg ctccacgctc gatccactgg cgagtgttag
     6061 taacagcact gttgcttcgt agcggagcat gacggccgtg ggaactcctc cttggtaaca
     6121 aggacccacg gggccaaaag ccacgcccac acgggcccgt catgtgtgca accccagcac
     6181 ggcgacttta ctgcgaaacc cactttaaag tgacattgaa actggtaccc acacactggt
     6241 gacaggctaa ggatgccctt caggtacccc gaggtaacac gcgacactcg ggatctgaga
     6301 aggggactgg ggcttctata aaagcgctcg gtttaaaaag cttctatgcc tgaataggtg
     6361 accggaggtc ggcacctttc ctttgcaatt actgacccta tgaatacact gactgtttga
     6421 caattaatca tcggcatagt atatcggcat agtataatac gactcactat aggagggcca
     6481 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     6541 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     6601 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6661 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6721 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6781 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6841 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6901 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6961 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     7021 acggcgagga tctcgtcgtg acacatggcg atgcctgctt gccgaatatc atggtggaaa
     7081 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     7141 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     7201 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     7261 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgaattcgct
     7321 aggattatcc ctaatacctg ccaccccact cttaatcagt ggtggaagaa cggtctcaga
     7381 actgtttgtt tcaattggcc atttaagttt agtagtaaaa gactggttaa tgataacaat
     7441 gcatcgtaaa accttcagaa ggaaaggaga atgttttgtg gaccactttg gttttctttt
     7501 ttgcgtgtgg cagttttaag ttattagttt ttaaaatcag tactttttaa tggaaacaac
     7561 ttgaccaaaa atttgtcaca gaattttgag acccattaaa aaagttaaat gagaaacctg
     7621 tgtgttcctt tggtcaacac cgagacattt aggtgaaaga catctaattc tggttttacg
     7681 aatctggaaa cttcttgaaa atgtaattct tgagttaaca cttctgggtg gagaataggg
     7741 ttgttttccc cccacataat tggaagggga aggaatatca tttaaagcta tgggagggtt
     7801 gctttgatta caacactgga gagaaatgca gcatgttgct gattgcctgt cactaaaaca
     7861 ggccaaaaac tgagtccttg ggttgcatag aaagctg
//
