LOCUS       sequence-80598-e        5900 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses YFP-PIF for red-light-induced binding to Phy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5900)
  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 5900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pAL175
FEATURES             Location/Qualifiers
     source          1..5900
                     /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(392..394,395..397,398..1108)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(437..458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(698..717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1045..1066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1452..1470)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1453..1470)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1490..1507)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1496..1720
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1766..2194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1853..1872)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2063..2084
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2203..2223)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2208..2538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2389..2524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2605..3399
                     /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(2659..2678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3269..3288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3573..3694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3610..3629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3664..3683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3743..3759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3743..3759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3756..3778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3767..3783
                     /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(3791..3797,3798..3815,3816..3821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3836..3857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3974..3991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4225..4244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4904..5695,5696..5764))
                     /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     5527..5546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5765..5869)
                     /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 catggtgagc aagggcgagg agctgttcac
      421 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      481 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      541 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
      601 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      661 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      721 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      781 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
      841 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
      901 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      961 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     1021 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1081 cactctcggc atggacgagc tgtacaaggg atccatgatg ttcttaccaa ccgattattg
     1141 ttgcaggtta agcgatcaag agtatatgga gcttgtgttt gagaatggcc agattcttgc
     1201 aaagggccaa agatccaacg tttctctgca taatcaacgt accaaatcga tcatggattt
     1261 gtatgaggca gagtataacg aggatttcat gaagagtatc atccatggtg gtggtggtgc
     1321 catcacaaat ctcggggaca cgcaggttgt tccacaaagt catgttgctg ctgcccatga
     1381 aacaaacatg ttggaaagca ataaacatgt tgactaagcg gccgctcgag catgcatcta
     1441 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     1501 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1561 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1621 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1681 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     1741 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1921 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1981 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2101 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     2161 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2221 ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat
     2281 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2341 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2401 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2461 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2521 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2581 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2641 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2701 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2761 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2821 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2881 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2941 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3001 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3061 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3121 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3181 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3241 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3301 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3361 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3421 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3481 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3541 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3601 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3661 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3721 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3781 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3841 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3901 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3961 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4861 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4921 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4981 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5041 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5101 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5161 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5221 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5281 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5341 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5401 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5461 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5521 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5581 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5641 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5701 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5761 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5821 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5881 gaaaagtgcc acctgacgtc
//
