LOCUS       sequence-112565-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Tiam DH-PH domains fused to mYFP-PIF6, for optogenetic 
            recruitment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  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 7091)
  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: pAL188
FEATURES             Location/Qualifiers
     source          1..7091
                     /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"
     promoter        169..379
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      238..373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(1583..1585,1586..1588,1589..2299)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1628..1649)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1889..1908)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2236..2257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2643..2661)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2644..2661)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2681..2698)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2687..2911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2957..3385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3044..3063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3254..3275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3394..3414)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3399..3729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3580..3715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3796..4590
                     /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(3850..3869)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4460..4479
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4764..4885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4801..4820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4855..4874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4934..4950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4934..4950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4947..4969)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4958..4974
                     /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(4982..4988,4989..5006,5007..5012))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5027..5048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5165..5182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5336..5924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5416..5435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6095..6886,6887..6955))
                     /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     6718..6737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6956..7060)
                     /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 caattgaggc aggcagaagt
      181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      301 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
      361 ttttttggag gcctaggctg gtaccatgcg acagctgtcg gatgcggata agctgcgcaa
      421 ggtgatctgt gaactgctgg agactgaacg cacctatgtg aaagacttaa actgcctcat
      481 ggagagatac ctgaagcccc ttcagaagga gacctttctc acccaggatg agcttgatgt
      541 actttttgga aatttaaccg aaatggtgga gtttcaagtc gagttcctta agactctaga
      601 agatggagta agactggtcc ctgacttgga aaagctggag aaggttgacc agttcaagaa
      661 agtgctcttc tctctggggg gatccttcct gtactacgcg gaccgcttca agctctacag
      721 tgccttctgt gcgagccaca caaaagtccc caaggtcctg gtgaaagcca agacggacac
      781 agccttcaag gcgttcctcg atgcccagaa cccgaggcag cagcactcat ccacgctgga
      841 gtcttacctc atcaagccca tccagagggt cctcaagtac ccgcttctgc tcagggagct
      901 gtttgcgctg accgatgcgg agagcgagga acactaccac ctggacgtgg ccatcaagac
      961 tatgaacaag gtcgccagtc acatcaatga gatgcagaag atccacgaag agtttggtgc
     1021 tgtgtttgac cagctgattg ctgagcagac gggagagaag aaagaggttg cagatctgag
     1081 catgggtgac ctgctcttgc acaccagcgt catctggctg aacccacccg cctcactggg
     1141 aaagtggaaa aaggagccag aattagcagc cttcgtcttc aaaacggccg tggtccttgt
     1201 atataaagac ggttccaagc agaagaagaa acttgttggc tctcacaggc tgtcaatcta
     1261 cgaggagtgg gaccctttcc ggtttcgcca catgatccct accgaagctt tgcaggtccg
     1321 agctctgccc agtgcagatg cagaggcaaa tgcggtatgc gaaattgtcc acgtgaaatc
     1381 agagtcagaa gggaggcccg agcgggtttt ccacctctgc tgcagttccc cagagagcag
     1441 aaaggacttt ctgaagtctg tgcattcgat cctgcgagat aaacacagaa gacagctcct
     1501 caaaacggaa gaattcagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg
     1561 tggtagtgct ggtggtgcta gcatggtgag caagggcgag gagctgttca ccggggtggt
     1621 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1681 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1741 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc
     1801 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1861 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1921 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1981 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2041 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2101 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2161 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     2221 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2281 catggacgag ctgtacaagg gatccatgat gttcttacca accgattatt gttgcaggtt
     2341 aagcgatcaa gagtatatgg agcttgtgtt tgagaatggc cagattcttg caaagggcca
     2401 aagatccaac gtttctctgc ataatcaacg taccaaatcg atcatggatt tgtatgaggc
     2461 agagtataac gaggatttca tgaagagtat catccatggt ggtggtggtg ccatcacaaa
     2521 tctcggggac acgcaggttg ttccacaaag tcatgttgct gctgcccatg aaacaaacat
     2581 gttggaaagc aataaacatg ttgactaagc ggccgctcga gcatgcatct agagggccct
     2641 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
     2701 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2761 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2821 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2881 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2941 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     3121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3301 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     3361 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3421 agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3661 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3721 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3781 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3841 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3901 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3961 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4021 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4081 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4141 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4201 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4261 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4321 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4381 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4441 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4501 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4561 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4621 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4681 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4741 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4801 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4861 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4921 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4981 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5041 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5101 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5161 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5221 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5281 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5341 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5401 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5461 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5521 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5581 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5641 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5701 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5761 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5821 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5881 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5941 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6001 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6061 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6121 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6181 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6241 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6301 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6361 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6421 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6481 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6541 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6601 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6661 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6721 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6781 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6841 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6901 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6961 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7021 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7081 cacctgacgt c
//
