LOCUS       Exported                6765 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses Ha-tagged VfAU1-LOV and myc-tagged casp9 (domains of 
            Opto-casp9) separated by P2A site .
ACCESSION   .
VERSION     .
KEYWORDS    VfAU1-LOV_P2A_casp9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6765)
  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 6765)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6765
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        158..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        538..741
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     692..712
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     786..805
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        786..804
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1280..1306
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1307..1363
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1364..1393
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1733..1744)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2279..2296)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2285..2509
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2555..2983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2642..2661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2852..2873
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2992..3012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2997..3326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3177..3312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3239..3258
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3393..4187
                     /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(3447..3466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4057..4076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4361..4482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4398..4417)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4452..4471
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4531..4547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4531..4547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4544..4566)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4555..4571
                     /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(4579..4609)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4624..4645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4762..4779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4933..5521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5013..5032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5692..6552)
                     /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     6315..6334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6553..6657)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6732..6751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
       61 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
      121 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
      181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      541 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      601 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      661 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      721 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
      781 gaaattaata cgactcacta tagggagacc caagctgacc atggccaccg gtcctgacta
      841 cagtctcgtg aaggctctgc aaatggcaca acagaatttt gtcattacag acgcctccct
      901 cccagacaac cctatcgtct acgccagtag agggtttctg acactgacag gctattctct
      961 cgaccagatc ctgggcagga actgcaggtt tctgcaaggg ccagaaacag acccaagagc
     1021 tgtggataag atcaggaatg ccatcaccaa aggcgttgat accagtgtct gtctgctgaa
     1081 ttatagacag gatggcacaa ccttctggaa tctcttcttc gtggctggac tcagagattc
     1141 taagggcaat attgtcaact acgtcggagt gcagtcaaag gtgagcgaag attatgccaa
     1201 gctgctggtc aacgagcaga acattgagta caaaggtgtg cgcaccagta acatgctgcg
     1261 cagaaagccc ggtgaattct acccatacga tgttccagat tacgctgcga ccaactttag
     1321 cctgctgaaa caggcgggcg atgtggaaga aaacccgggc ccggaacaaa aactcatctc
     1381 agaagaggat ctgggatccc ccgggggatt tggtgatgtc ggtgctcttg agagtttgag
     1441 gggaaatgca gatttggctt acatcctgag catggagccc tgtggccact gcctcattat
     1501 caacaatgtg aacttctgcc gtgagtccgg gctccgcacc cgcactggct ccaacatcga
     1561 ctgtgagaag ttgcggcgtc gcttctcctc gctgcatttc atggtggagg tgaagggcga
     1621 cctgactgcc aagaaaatgg tgctggcttt gctggagctg gcgcggcagg accacggtgc
     1681 tctggactgc tgcgtggtgg tcattctctc tcacggctgt caggccagcc acctgcagtt
     1741 cccaggggct gtctacggca cagatggatg ccctgtgtcg gtcgagaaga ttgtgaacat
     1801 cttcaatggg accagctgcc ccagcctggg agggaagccc aagctctttt tcatccaggc
     1861 ctgtggtggg gagcagaaag accatgggtt tgaggtggcc tccacttccc ctgaagacga
     1921 gtcccctggc agtaaccccg agccagatgc caccccgttc caggaaggtt tgaggacctt
     1981 cgaccagctg gacgccatat ctagtttgcc cacacccagt gacatctttg tgtcctactc
     2041 tactttccca ggttttgttt cctggaggga ccccaagagt ggctcctggt acgttgagac
     2101 cctggacgac atctttgagc agtgggctca ctctgaagac ctgcagtccc tcctgcttag
     2161 ggtcgctaat gctgtttcgg tgaaagggat ttataaacag atgcctggtt gctttaattt
     2221 cctccggaaa aaacttttct ttaaaacatc acccgggtaa gtttaaaccg ctgatcagcc
     2281 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2341 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2401 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2461 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2521 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2581 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2641 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2701 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2761 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2821 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2881 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2941 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3001 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3061 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3121 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3181 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3241 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3301 tttttggagg cctaggcttt tgcaaaaagc tctcgggagc ttgtatatcc attttcggat
     3361 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3421 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3481 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3541 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3601 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3661 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3721 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3781 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3841 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3901 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3961 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4021 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4081 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4141 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4201 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4261 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4321 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4381 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4441 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4501 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4561 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4621 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4681 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4741 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4801 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4861 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4921 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4981 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5041 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5101 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5161 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5221 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5281 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5341 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5401 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5461 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5521 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5581 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5641 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5701 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5761 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5821 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5881 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5941 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6001 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6061 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6121 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6181 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6241 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6301 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6361 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6421 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6481 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6541 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6601 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6661 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6721 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccag
//
