LOCUS       sequence-10328-e        6742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N terminal domain of PIF3 fused to mYFP, for 
            red-light-induced binding to Phy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6742)
  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 6742)
  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: pAL130
FEATURES             Location/Qualifiers
     source          1..6742
                     /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(1540..1542,1543..1545,1546..2259)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1585..1606)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1846..1865)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2193..2214
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2294..2312)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2295..2312)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2332..2349)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2338..2562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2608..3036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2695..2714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2905..2926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3045..3065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3050..3380
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3231..3366
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3293..3312
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3447..4241
                     /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(3501..3520)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4111..4130
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4415..4536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4452..4471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4506..4525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4585..4601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4585..4601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4598..4620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4609..4625
                     /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(4633..4639,4640..4657,4658..4663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4678..4699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4816..4833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4987..5575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5067..5086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5746..6537,6538..6606))
                     /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     6369..6388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6607..6711)
                     /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 atgcctctgt ttgagctttt caggctcacc aaagctaagc ttgaatctgc tcaagacagg
      961 aacccttctc cacctgtaga tgaagttgtg gagctggtgt gggaaaatgg tcagatatca
     1021 actcaaagtc agtcaagtag atcgaggaac attcctccac cacaagcaaa ctcttctaga
     1081 gctagagaga ttggaaatgg ctcaaagacg actatggtgg acgagatccc tatgtcagtg
     1141 ccatcactaa tgacgggttt gagtcaagac gatgactttg ttccatggtt gaatcatcat
     1201 ccctcccttg atggatattg ctctgatttc ttgcgtgatg tgtcgtctcc tgttactgtc
     1261 aacgagcaag agagtgatat ggcggtaaac caaactgctt tcccgttgtt tcagagaaga
     1321 aaggatggca atgaatcagc tcctgctgct tcttcgtcgc agtataacgg tttccaatcg
     1381 cattctctgt atggaagtga tagagctaga gatcttccta gccaacaaac caatccggat
     1441 cggtttactc agacgcagga accactaatt actagtaaca agcctagttt ggtcaacttt
     1501 tcacatttct tacgccctgc aacttttgcg aagggatcca tggtgagcaa gggcgaggag
     1561 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1621 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1681 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     1741 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1801 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1861 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1921 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1981 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2041 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2101 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     2161 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2221 gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgctcg agcatgcatc
     2281 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2341 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2401 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2461 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2521 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2581 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2641 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2701 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2761 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2821 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2881 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2941 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     3001 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3061 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     3121 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3181 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3241 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3301 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3361 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3421 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3481 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3541 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3601 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3661 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3721 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3781 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3841 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3901 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3961 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4021 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4081 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4141 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4201 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4261 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4321 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4381 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4441 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4501 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4561 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4621 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4681 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4741 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5161 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5401 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5821 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5881 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5941 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6181 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6241 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6421 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6601 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6661 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6721 ccgaaaagtg ccacctgacg tc
//
