LOCUS       sequence-257061-        6660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Opto-casp9_casp9-VfAU1-LOV (light-activated caspase9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6660)
  AUTHORS   Tichy AM, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak H
  TITLE     Engineering Strategy and Vector Library for the Rapid Generation of 
            Modular Light-Controlled Protein-Protein Interactions.
  JOURNAL   J Mol Biol. 2019 May 29. pii: S0022-2836(19)30317-1. doi: 
            10.1016/j.jmb.2019.05.033.
  PUBMED    31150735
REFERENCE   2  (bases 1 to 6660)
  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: "J Mol Biol.
            2019 May 29. pii: S0022-2836(19)30317-1. doi: 
            10.1016/j.jmb.2019.05.033."
COMMENT     Sequence Label: Opto-casp9_casp9-VfAU1-LOV
FEATURES             Location/Qualifiers
     source          1..6660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        143..522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..726
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     677..697
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     771..790
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        771..789
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1145..1156)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2159..2176)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2165..2389
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2435..2863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2522..2541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2732..2753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2872..2892)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2877..3206
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3057..3192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3119..3138
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3273..4067
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3327..3346)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3937..3956
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4241..4362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4278..4297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4332..4351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4411..4427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4411..4427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4424..4446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4435..4451
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4459..4465,4466..4483,4484..4489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4504..4525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4642..4659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4813..5401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5572..6363,6364..6432))
                     /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     6195..6214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6433..6537)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6612..6631)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
       61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      721 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      781 cactataggg agacccaagc tgaccatggc caccggggga tttggtgatg tcggtgctct
      841 tgagagtttg aggggaaatg cagatttggc ttacatcctg agcatggagc cctgtggcca
      901 ctgcctcatt atcaacaatg tgaacttctg ccgtgagtcc gggctccgca cccgcactgg
      961 ctccaacatc gactgtgaga agttgcggcg tcgcttctcc tcgctgcatt tcatggtgga
     1021 ggtgaagggc gacctgactg ccaagaaaat ggtgctggct ttgctggagc tggcgcggca
     1081 ggaccacggt gctctggact gctgcgtggt ggtcattctc tctcacggct gtcaggccag
     1141 ccacctgcag ttcccagggg ctgtctacgg cacagatgga tgccctgtgt cggtcgagaa
     1201 gattgtgaac atcttcaatg ggaccagctg ccccagcctg ggagggaagc ccaagctctt
     1261 tttcatccag gcctgtggtg gggagcagaa agaccatggg tttgaggtgg cctccacttc
     1321 ccctgaagac gagtcccctg gcagtaaccc cgagccagat gccaccccgt tccaggaagg
     1381 tttgaggacc ttcgaccagc tggacgccat atctagtttg cccacaccca gtgacatctt
     1441 tgtgtcctac tctactttcc caggttttgt ttcctggagg gaccccaaga gtggctcctg
     1501 gtacgttgag accctggacg acatctttga gcagtgggct cactctgaag acctgcagtc
     1561 cctcctgctt agggtcgcta atgctgtttc ggtgaaaggg atttataaac agatgcctgg
     1621 ttgctttaat ttcctccgga aaaaactttt ctttaaaaca tcacccggtg aattctcagg
     1681 ctctggatcc cccggtcctg actacagtct cgtgaaggct ctgcaaatgg cacaacagaa
     1741 ttttgtcatt acagacgcct ccctcccaga caaccctatc gtctacgcca gtagagggtt
     1801 tctgacactg acaggctatt ctctcgacca gatcctgggc aggaactgca ggtttctgca
     1861 agggccagaa acagacccaa gagctgtgga taagatcagg aatgccatca ccaaaggcgt
     1921 tgataccagt gtctgtctgc tgaattatag acaggatggc acaaccttct ggaatctctt
     1981 cttcgtggct ggactcagag attctaaggg caatattgtc aactacgtcg gagtgcagtc
     2041 aaaggtgagc gaagattatg ccaagctgct ggtcaacgag cagaacattg agtacaaagg
     2101 tgtgcgcacc agtaacatgc tgcgcagaaa gcccgggtaa gtttaaaccg ctgatcagcc
     2161 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2221 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2281 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2341 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2401 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2641 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2761 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2821 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2881 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2941 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3001 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3061 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3121 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3181 tttttggagg cctaggcttt tgcaaaaagc tctcgggagc ttgtatatcc attttcggat
     3241 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3301 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3361 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3421 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3481 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3541 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3601 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3661 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3721 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3781 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3841 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3901 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3961 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4021 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4081 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4141 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4201 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4261 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4321 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4381 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4441 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4501 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4561 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4621 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4681 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5221 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5641 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5761 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6001 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6061 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6241 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6421 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6481 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6541 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6601 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
//
