LOCUS       Exported                7716 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses Opto-casp9_AtCRY2-PHR-casp9 (light-activated caspase9).
ACCESSION   .
VERSION     .
KEYWORDS    Opto-casp9_AtCRY2-PHR-casp9
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 May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7716
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        156..535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        536..739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     690..710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     784..803
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        784..802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2682..2693)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3228..3245)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3234..3458
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3504..3932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3591..3610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3801..3822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3941..3961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3946..4275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4126..4261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4188..4207
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4342..5136
                     /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(4396..4415)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5006..5025
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5310..5431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5347..5366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5401..5420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5480..5496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5480..5496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5493..5515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5504..5520
                     /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(5528..5558)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5573..5594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5711..5728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5882..6470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5962..5981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6641..7501)
                     /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     7264..7283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7502..7606)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7681..7700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
       61 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      121 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      541 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      601 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      661 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      721 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      781 aattaatacg actcactata gggagaccca agctgaccat ggccaccggt aagatggaca
      841 agaaaaccat agtctggttt cgccgcgatt tgaggataga ggataaccct gcgttggccg
      901 cagcggccca cgaaggcagc gtgttccccg tgttcatatg gtgcccagag gaggaaggcc
      961 agttctaccc aggtcgcgct agtcgctggt ggatgaaaca gtctctcgca catctctccc
     1021 aatctttgaa agctctcggg tctgacctta cgctgattaa gacccacaat actattagtg
     1081 caatcttgga ctgcatccgc gttaccggcg cgaccaaagt ggtgtttaat catctgtatg
     1141 acccggtaag tctggtacgc gaccatacag ttaaggagaa gttggttgaa aggggaattt
     1201 cagtacagag ttataatgga gacctcctct acgaaccttg ggaaatatac tgtgagaagg
     1261 gaaagccatt tacatcattt aactcatact ggaaaaagtg tcttgacatg agcatagagt
     1321 ctgtcatgtt gcccccgccg tggcggctta tgcccatcac cgcggccgcc gaagctatct
     1381 gggcgtgtag tattgaagaa cttggcttgg agaacgaagc agaaaaacct agcaatgcac
     1441 tgttgacgcg cgcctggtcc cctgggtgga gtaacgcaga taaattgctt aacgagttca
     1501 tcgaaaagca acttatcgac tacgctaaga atagcaaaaa agtggtgggc aattcaacct
     1561 ccctcctgtc tccgtacttg catttcggcg agattagcgt gcggcacgtc ttccaatgcg
     1621 cccgaatgaa acaaataatc tgggcgcggg ataaaaacag tgagggtgaa gaaagcgcag
     1681 acttgttcct ccgaggaatc ggtctgcgag aatatagtcg ctacatttgt ttcaattttc
     1741 cctttacgca tgagcagagc ctccttagtc acttgcgatt ctttccttgg gacgcagatg
     1801 ttgacaaatt taaagcatgg cggcaaggta gaacaggcta cccattggta gatgcgggta
     1861 tgcgagaact ctgggccacg gggtggatgc ataaccgaat cagggtaata gtaagtagtt
     1921 tcgcagttaa gtttcttttg cttccatgga agtgggggat gaagtatttc tgggacactt
     1981 tgctcgatgc ggatcttgaa tgcgatatat tgggttggca atatatttcc gggtcaatcc
     2041 ctgacggcca tgagctggac agattggaca atcctgcgct ccagggtgcg aaatacgatc
     2101 ccgaaggtga atacattaga caatggcttc cagaacttgc caggttgccc acggaatgga
     2161 ttcaccaccc atgggacgcc cctcttacgg ttttgaaggc gagtggtgtg gagctcggta
     2221 ccaattacgc aaaaccgatt gttgacattg ataccgcgcg ggagttgctg gctaaagcca
     2281 tttcacgaac ccggaaagcc caaatcatga ttggtgctgc aaccggtgaa ttctcaggct
     2341 ctggatcccc cgggggattt ggtgatgtcg gtgctcttga gagtttgagg ggaaatgcag
     2401 atttggctta catcctgagc atggagccct gtggccactg cctcattatc aacaatgtga
     2461 acttctgccg tgagtccggg ctccgcaccc gcactggctc caacatcgac tgtgagaagt
     2521 tgcggcgtcg cttctcctcg ctgcatttca tggtggaggt gaagggcgac ctgactgcca
     2581 agaaaatggt gctggctttg ctggagctgg cgcggcagga ccacggtgct ctggactgct
     2641 gcgtggtggt cattctctct cacggctgtc aggccagcca cctgcagttc ccaggggctg
     2701 tctacggcac agatggatgc cctgtgtcgg tcgagaagat tgtgaacatc ttcaatggga
     2761 ccagctgccc cagcctggga gggaagccca agctcttttt catccaggcc tgtggtgggg
     2821 agcagaaaga ccatgggttt gaggtggcct ccacttcccc tgaagacgag tcccctggca
     2881 gtaaccccga gccagatgcc accccgttcc aggaaggttt gaggaccttc gaccagctgg
     2941 acgccatatc tagtttgccc acacccagtg acatctttgt gtcctactct actttcccag
     3001 gttttgtttc ctggagggac cccaagagtg gctcctggta cgttgagacc ctggacgaca
     3061 tctttgagca gtgggctcac tctgaagacc tgcagtccct cctgcttagg gtcgctaatg
     3121 ctgtttcggt gaaagggatt tataaacaga tgcctggttg ctttaatttc ctccggaaaa
     3181 aacttttctt taaaacatca cccgggtaag tttaaaccgc tgatcagcct cgactgtgcc
     3241 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3301 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3361 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3421 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3481 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3841 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3901 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4021 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4081 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4141 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4201 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4261 ctaggctttt gcaaaaagct ctcgggagct tgtatatcca ttttcggatc tgatcaagag
     4321 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4381 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4441 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4501 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4561 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4621 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4681 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4741 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4801 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4861 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4921 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4981 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5041 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5101 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5161 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5221 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5281 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5341 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5401 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5461 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5521 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5581 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5641 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5701 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5761 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5821 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5881 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5941 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6001 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6061 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6121 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6181 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6241 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6301 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6361 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6421 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6481 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6541 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6601 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6661 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6721 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6781 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6841 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6901 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6961 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7021 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7081 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7141 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7201 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7261 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7321 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7381 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7441 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7501 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7561 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7621 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     7681 tacaatctgc tctgatgccg catagttaag ccagta
//
