LOCUS       sequence-257059-        7716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Opto-casp9_casp9-AtCRY2-PHR (light-activated caspase9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7716)
  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 7716)
  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-AtCRY2-PHR
FEATURES             Location/Qualifiers
     source          1..7716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        136..515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        516..719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     670..690
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     764..783
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        764..782
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1138..1149)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3208..3225)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3214..3438
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3484..3912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3571..3590)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3781..3802
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3921..3941)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3926..4255
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4106..4241
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4168..4187
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4322..5116
                     /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(4376..4395)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4986..5005
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5290..5411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5327..5346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5381..5400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5460..5476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5460..5476)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5473..5495)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5484..5500
                     /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(5508..5514,5515..5532,5533..5538))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5553..5574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5691..5708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5862..6450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5942..5961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6621..7412,7413..7481))
                     /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     7244..7263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7482..7586)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7661..7680)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
       61 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      121 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
      181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      481 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      541 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      601 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      661 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      721 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
      781 gggagaccca agctgaccat ggccaccggg ggatttggtg atgtcggtgc tcttgagagt
      841 ttgaggggaa atgcagattt ggcttacatc ctgagcatgg agccctgtgg ccactgcctc
      901 attatcaaca atgtgaactt ctgccgtgag tccgggctcc gcacccgcac tggctccaac
      961 atcgactgtg agaagttgcg gcgtcgcttc tcctcgctgc atttcatggt ggaggtgaag
     1021 ggcgacctga ctgccaagaa aatggtgctg gctttgctgg agctggcgcg gcaggaccac
     1081 ggtgctctgg actgctgcgt ggtggtcatt ctctctcacg gctgtcaggc cagccacctg
     1141 cagttcccag gggctgtcta cggcacagat ggatgccctg tgtcggtcga gaagattgtg
     1201 aacatcttca atgggaccag ctgccccagc ctgggaggga agcccaagct ctttttcatc
     1261 caggcctgtg gtggggagca gaaagaccat gggtttgagg tggcctccac ttcccctgaa
     1321 gacgagtccc ctggcagtaa ccccgagcca gatgccaccc cgttccagga aggtttgagg
     1381 accttcgacc agctggacgc catatctagt ttgcccacac ccagtgacat ctttgtgtcc
     1441 tactctactt tcccaggttt tgtttcctgg agggacccca agagtggctc ctggtacgtt
     1501 gagaccctgg acgacatctt tgagcagtgg gctcactctg aagacctgca gtccctcctg
     1561 cttagggtcg ctaatgctgt ttcggtgaaa gggatttata aacagatgcc tggttgcttt
     1621 aatttcctcc ggaaaaaact tttctttaaa acatcacccg gtgaattctc aggctctgga
     1681 tcccccggta agatggacaa gaaaaccata gtctggtttc gccgcgattt gaggatagag
     1741 gataaccctg cgttggccgc agcggcccac gaaggcagcg tgttccccgt gttcatatgg
     1801 tgcccagagg aggaaggcca gttctaccca ggtcgcgcta gtcgctggtg gatgaaacag
     1861 tctctcgcac atctctccca atctttgaaa gctctcgggt ctgaccttac gctgattaag
     1921 acccacaata ctattagtgc aatcttggac tgcatccgcg ttaccggcgc gaccaaagtg
     1981 gtgtttaatc atctgtatga cccggtaagt ctggtacgcg accatacagt taaggagaag
     2041 ttggttgaaa ggggaatttc agtacagagt tataatggag acctcctcta cgaaccttgg
     2101 gaaatatact gtgagaaggg aaagccattt acatcattta actcatactg gaaaaagtgt
     2161 cttgacatga gcatagagtc tgtcatgttg cccccgccgt ggcggcttat gcccatcacc
     2221 gcggccgccg aagctatctg ggcgtgtagt attgaagaac ttggcttgga gaacgaagca
     2281 gaaaaaccta gcaatgcact gttgacgcgc gcctggtccc ctgggtggag taacgcagat
     2341 aaattgctta acgagttcat cgaaaagcaa cttatcgact acgctaagaa tagcaaaaaa
     2401 gtggtgggca attcaacctc cctcctgtct ccgtacttgc atttcggcga gattagcgtg
     2461 cggcacgtct tccaatgcgc ccgaatgaaa caaataatct gggcgcggga taaaaacagt
     2521 gagggtgaag aaagcgcaga cttgttcctc cgaggaatcg gtctgcgaga atatagtcgc
     2581 tacatttgtt tcaattttcc ctttacgcat gagcagagcc tccttagtca cttgcgattc
     2641 tttccttggg acgcagatgt tgacaaattt aaagcatggc ggcaaggtag aacaggctac
     2701 ccattggtag atgcgggtat gcgagaactc tgggccacgg ggtggatgca taaccgaatc
     2761 agggtaatag taagtagttt cgcagttaag tttcttttgc ttccatggaa gtgggggatg
     2821 aagtatttct gggacacttt gctcgatgcg gatcttgaat gcgatatatt gggttggcaa
     2881 tatatttccg ggtcaatccc tgacggccat gagctggaca gattggacaa tcctgcgctc
     2941 cagggtgcga aatacgatcc cgaaggtgaa tacattagac aatggcttcc agaacttgcc
     3001 aggttgccca cggaatggat tcaccaccca tgggacgccc ctcttacggt tttgaaggcg
     3061 agtggtgtgg agctcggtac caattacgca aaaccgattg ttgacattga taccgcgcgg
     3121 gagttgctgg ctaaagccat ttcacgaacc cggaaagccc aaatcatgat tggtgctgca
     3181 accgggtaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3241 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3301 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3361 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3421 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3481 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3541 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3661 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3781 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3841 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3901 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3961 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4081 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4141 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4201 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4261 ctcgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4321 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4381 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4441 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4501 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4561 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4621 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4681 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4741 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4801 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4861 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4921 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4981 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5041 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5101 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5161 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5221 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5281 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5401 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6301 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6721 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6901 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7561 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7621 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     7681 catagttaag ccagtatctg ctccctgctt gtgtgt
//
