LOCUS       sequence-10325-e        8050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal domains of PhyB (with Y276H mutation), fused to
            mCFP and HRas C terminal CAAX palmitoylation sequence. For 
            red-light-induced binding to PIF6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8050)
  AUTHORS   Levskaya A, Weiner OD, Lim WA, Voigt CA
  TITLE     Spatiotemporal control of cell signalling using a light-switchable 
            protein interaction.
  JOURNAL   Nature. 2009 Oct 15;461(7266):997-1001.
  PUBMED    19749742
REFERENCE   2  (bases 1 to 8050)
  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: "Nature."; 
            date: "2009-10-15"; volume: "461(7266)"; pages: "997-1001"
COMMENT     Sequence Label: pAL113
FEATURES             Location/Qualifiers
     source          1..8050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2812..2814,2815..2817,2818..3528)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2857..2878)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3118..3137)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3465..3486
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3602..3620)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3603..3620)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3640..3657)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3646..3870
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3916..4344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4003..4022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4213..4234
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4353..4373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4358..4688
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4539..4674
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4601..4620
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4755..5549
                     /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(4809..4828)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5419..5438
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5723..5844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5760..5779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5814..5833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5893..5909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5893..5909)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5906..5928)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5917..5933
                     /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(5941..5947,5948..5965,5966..5971))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5986..6007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6124..6141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6295..6883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6375..6394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7054..7845,7846..7914))
                     /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     7677..7696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7915..8019)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 atggtttccg gagtcggggg tagtggcggt ggccgtggcg gtggccgtgg cggagaagaa
      961 gaaccgtcgt caagtcacac tcctaataac cgaagaggag gagaacaagc tcaatcgtcg
     1021 ggaacgaaat ctctcagacc aagaagcaac actgaatcaa tgagcaaagc aattcaacag
     1081 tacaccgtcg acgcaagact ccacgccgtt ttcgaacaat ccggcgaatc agggaaatca
     1141 ttcgactact cacaatcact caaaacgacg acgtacggtt cctctgtacc tgagcaacag
     1201 atcacagctt atctctctcg aatccagcga ggtggttaca ttcagccttt cggatgtatg
     1261 atcgccgtcg atgaatccag tttccggatc atcggttaca gtgaaaacgc cagagaaatg
     1321 ttagggatta tgcctcaatc tgttcctact cttgagaaac ctgagattct agctatggga
     1381 actgatgtga gatctttgtt cacttcttcg agctcgattc tactcgagcg tgctttcgtt
     1441 gctcgagaga ttaccttgtt aaatccggtt tggatccatt ccaagaatac tggtaaaccg
     1501 ttttacgcca ttcttcatag gattgatgtt ggtgttgtta ttgatttaga gccagctaga
     1561 actgaagatc ctgcgctttc tattgctggt gctgttcaat cgcagaaact cgcggttcgt
     1621 gcgatttctc agttacaggc tcttcctggt ggagatatta agcttttgtg tgacactgtc
     1681 gtggaaagtg tgagggactt gactggttat gatcgtgtta tggttcataa gtttcatgaa
     1741 gatgagcatg gagaagttgt agctgagagt aaacgagatg atttagagcc ttatattgga
     1801 ctgcattatc ctgctactga tattcctcaa gcgtcaaggt tcttgtttaa gcagaaccgt
     1861 gtccgaatga tagtagattg caatgccaca cctgttcttg tggtccagga cgataggcta
     1921 actcagtcta tgtgcttggt tggttctact cttagggctc ctcatggttg tcactctcag
     1981 tatatggcta acatgggatc tattgcgtct ttagcaatgg cggttataat caatggaaat
     2041 gaagatgatg ggagcaatgt agctagtgga agaagctcga tgaggctttg gggtttggtt
     2101 gtttgccatc acacttcttc tcgctgcata ccgtttccgc taaggtatgc ttgtgagttt
     2161 ttgatgcagg ctttcggttt acagttaaac atggaattgc agttagcttt gcaaatgtca
     2221 gagaaacgcg ttttgagaac gcagacactg ttatgtgata tgcttctgcg tgactcgcct
     2281 gctggaattg ttacacagag tcccagtatc atggacttag tgaaatgtga cggtgcagca
     2341 tttctttacc acgggaagta ttacccgttg ggtgttgctc ctagtgaagt tcagataaaa
     2401 gatgttgtgg agtggttgct tgcgaatcat gcggattcaa ccggattaag cactgatagt
     2461 ttaggcgatg cggggtatcc cggtgcagct gcgttagggg atgctgtgtg cggtatggca
     2521 gttgcatata tcacaaaaag agactttctt ttttggtttc gatctcacac tgcgaaagaa
     2581 atcaaatggg gaggcgctaa gcatcatccg gaggataaag atgatgggca acgaatgcat
     2641 cctcgttcgt cctttcaggc ttttcttgaa gttgttaaga gccggagtca gccatgggaa
     2701 actgcggaaa tggatgcgat tcactcgctc cagcttattc tgagagactc ttttaaagaa
     2761 tctgaggcgg aattcgatag tgctggtagt gctggtagtg ctggtactag tatggtgagc
     2821 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2881 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     2941 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3001 accctgacct ggggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3061 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3121 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3181 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3241 tacaactaca tcagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     3301 gccaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3361 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3421 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3481 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagct gcagggctgc
     3541 atgagctgca agtgtgtgct ctcctaagcg gccgctcgag catgcatcta gagggcccta
     3601 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3661 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3721 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3781 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3841 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3901 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3961 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4021 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     4081 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4141 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4201 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4261 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     4321 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4381 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4441 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4501 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4561 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4621 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4681 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4741 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4801 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4861 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4921 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4981 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5041 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5101 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5161 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5221 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5281 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5341 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5401 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5461 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5521 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5581 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5641 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5701 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5761 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5821 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5881 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5941 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6001 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6061 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6481 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6721 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7021 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7141 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7201 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7981 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8041 acctgacgtc
//
