LOCUS       sequence-80601-e        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Tim 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 7061)
  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 7061)
  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: pAL190
FEATURES             Location/Qualifiers
     source          1..7061
                     /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(1553..1555,1556..1558,1559..2269)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1598..1619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1859..1878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2206..2227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2613..2631)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2614..2631)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2651..2668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2657..2881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2927..3355
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3014..3033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3224..3245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3364..3384)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3369..3699
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3550..3685
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3766..4560
                     /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(3820..3839)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4430..4449
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4734..4855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4771..4790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4825..4844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4904..4920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4904..4920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4917..4939)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4928..4944
                     /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(4952..4958,4959..4976,4977..4982))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4997..5018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5135..5152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5306..5894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5386..5405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6065..6856,6857..6925))
                     /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     6688..6707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6926..7030)
                     /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 cgcctctgcc tctgagctat tccagaagta gtgaggaggc
      361 ttttttggag gcctaggcta agcttggtac catgctctcc atgacccatg aagaccaaaa
      421 gctgcaagag gtcaaatttg agctgattgt gtcagaggcc tcctacctgc gcagtctaaa
      481 catagctgtg gatcatttcc aactttcaac ttcactccgg gccacacttt ccaaccagga
      541 gcaccaatgg ctcttctctc gtttacagga tgtgcgagac gtcagcgcca cgttcctttc
      601 agacctggaa gagaactttg agaacaatat cttctccttc caagtatgtg acgtagtcct
      661 gaaccacgcc ccagacttcc gccgggtcta cctgccttat gtcaccaacc agacctatca
      721 ggaacgcacc ttccagagcc tgatgaatag caacagcaat ttccgggagg tcttggagaa
      781 gctggagagc gaccccgtct gccagcgcct ttccctcaag tcctttctga ttctgccctt
      841 ccaacgcatc acccgcctca aactgctgct ccagaacatt ctgaagagaa cacagcctgg
      901 ctcctcggag gaggcagagg ccacgaaggc acaccacgcc ctggagcagc tgatccggga
      961 ctgcaataac aatgtccaga gtatgcgacg gacagaggag ctaatctacc tgagccagaa
     1021 gattgagttt gagtgcaaaa tattcccgct catttctcag tcacgctggc tggtgaaaag
     1081 tggggagctg acagccttgg agttcagtgc ttccccaggg ctacgaagga agctgaacac
     1141 gcgtccagtc cacctgcacc tcttcaatga ctgtctgctg ctgtctcggc cccgagaggg
     1201 tagccgattc ctggtatttg accatgctcc cttctcctcc attcgggggg aaaagtgtga
     1261 aatgaagcta catggacctc acaaaaacct gttccgactc tttctgcggc agaacactca
     1321 gggcgcccag gccgagttcc tcttccgcac ggagactcaa agtgaaaagc ttcggtggat
     1381 ctcagccttg gccatgccaa gagaggagtt ggaccttctg gagtgttaca actcccccca
     1441 ggtacagtgc cttcgagcct acaagccccg agagaatgat gaattcagtg ctggtggtag
     1501 tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggtgcta gcatggtgag
     1561 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1621 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1681 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1741 caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1801 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1861 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1921 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1981 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2041 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2101 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2161 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2221 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gatccatgat
     2281 gttcttacca accgattatt gttgcaggtt aagcgatcaa gagtatatgg agcttgtgtt
     2341 tgagaatggc cagattcttg caaagggcca aagatccaac gtttctctgc ataatcaacg
     2401 taccaaatcg atcatggatt tgtatgaggc agagtataac gaggatttca tgaagagtat
     2461 catccatggt ggtggtggtg ccatcacaaa tctcggggac acgcaggttg ttccacaaag
     2521 tcatgttgct gctgcccatg aaacaaacat gttggaaagc aataaacatg ttgactaagc
     2581 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     2641 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2701 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2761 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2821 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2881 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2941 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3001 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3061 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     3121 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3181 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3241 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     3301 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3361 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccaggc aggcagaagt
     3421 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3481 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3541 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3601 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3661 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3721 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3781 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3841 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3901 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3961 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4021 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4081 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4141 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4201 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4261 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4321 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4381 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4441 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4501 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4561 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4621 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4681 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4741 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4801 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4861 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4921 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4981 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5041 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5101 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5161 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5221 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5281 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5341 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5401 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5461 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     5521 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5581 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5641 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5701 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5761 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5821 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5881 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5941 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6001 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6061 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6121 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6181 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6241 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6301 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6361 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6421 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6481 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6541 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6601 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6661 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6721 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6781 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6841 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6901 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6961 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7021 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
