LOCUS       sequence-328895-        7137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For light-inducible Cre recombinase (LiCre) expression under the 
            Met17 promoter in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  AUTHORS   Duplus-Bottin H, Spichty M, Triqueneaux G, Place C, Mangeot PE, 
            Ohlmann T, Vittoz F, Yvert G
  TITLE     A single-chain and fast-responding light-inducible Cre recombinase 
            as a novel optogenetic switch.
  JOURNAL   Elife. 2021 Feb 23;10. pii: 61268. doi: 10.7554/eLife.61268.
  PUBMED    33620312
REFERENCE   2  (bases 1 to 7137)
  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: "Elife."; 
            date: "2021-02-23"; volume: "10. pii: 61268. doi"; pages: " 
            10.7554/eLife.61268"
COMMENT     Sequence Label: pGY466
FEATURES             Location/Qualifiers
     source          1..7137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    69..572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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     complement(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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)."
     primer_bind     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2754..2774
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2825..3174
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3175..3191
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3175..3191
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3225..3241)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3226..3242)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             3256..3273
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4983..5230
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4999..5017)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5248..5267)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5249..5267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5277..5294)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5277..5293)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5286..5308)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5434..5889
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5521..5540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5731..5752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5992..6666)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(6667..6947)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6968..6987)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7087..7109
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
      121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
      181 aataaagaag gtagtagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
      241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
      301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
      361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
      421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
      481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
      541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2761 accctcacta aagggaacaa aagctggagc tccggatgca agggttcgaa tcccttagct
     2821 ctcattattt tttgcttttt ctcttgaggt cacatgatcg caaaatggca aatggcacgt
     2881 gaagctgtcg atattgggga actgtggtgg ttggctaaaa ccaaattaag ttagtcaagg
     2941 cgccatcctc ataaaaactg tgtaacataa taaccgaagt gtcgaaaagg tggcaccttg
     3001 tccaattgaa cacgctcgat gaaaaaaata agatatatat aaggttaagt aaagcgtctg
     3061 ttagaaagga agtttttcct ttttcttgct ctcttgtctt ttcatctact atttccttcg
     3121 tgtaatacag ggtcgtcaga tacatagata caattctatt acccccatcc atactctaga
     3181 actagtggat cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc
     3241 gacatgagag gttcacatca tcaccaccat cacggttctt tagcaacaac tctggaaaga
     3301 atcgagaaaa acttcgtcat cacggacccg aggctacccg acaaccctat cattttcgca
     3361 tccgactctt ttttacaatt aactgaatat tctagagaag aaattctggg ccgtaattgt
     3421 aggtttttac agggtccaga aacagataga gcaacagtta gaaagattag agatgctatt
     3481 gataatcaga ctgaggtaac agttcaacta ataaactata ctaaatccgg gaagaaattt
     3541 tggaatgtct tccacttaca gccaatgaga gattacaaag gtgatgtcca gtattttatc
     3601 ggagttcagt tagacggcac agaacgtctg cacggtgccg cggagaggga agctgtttgt
     3661 cttattaaga aaaccgcatt tcaaatcgac agtgatgagg ttcgcaagaa cctgatggac
     3721 atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct gtccgtttgc
     3781 cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc agaacctgaa
     3841 gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa aactatccag
     3901 caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg accaagtgac
     3961 agcaatgctg tttcactggt tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa
     4021 cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg ttcactcatg
     4081 gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat tgcttataac
     4141 accctgttac gtatagccga aattgccagg atcagggtta aagatatctc acgtactgac
     4201 ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac cgcaggtgta
     4261 gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc cgtctctggt
     4321 gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg
     4381 ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga agcaactcat
     4441 cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg gtctggacac
     4501 agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat accggagatc
     4561 atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg taacctggat
     4621 agtgaaacag gggcaatggt gcgcctgctg gcagctggcg attagccatt aacgcgtaaa
     4681 tgattgctat aattatttga tatttatggt gacatatgag aaaggatttc aacatcgacg
     4741 gaaaatatgt agtgctgtct gtaagcacta atattcagtc gccagccgtc attgtcactg
     4801 taaagctgag cgatagaatg cctgatattg actcaatatc cgttgcgttt cctgtcaaaa
     4861 gtatgcgtag tgctgaacat ttcgtgatga atgccaccga ggaagaagca cggcgcggtt
     4921 ttgctaaagt gatgtctgag tttggcgaac tcttgggtaa ggttggaatt gtcgacctcg
     4981 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     5041 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     5101 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     5161 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     5221 tttaatttgc ggccggtacc caattcgccc tatagtgagt cgtattaggc gcgctcactg
     5281 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5341 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5401 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     5461 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5521 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5581 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5641 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5701 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5761 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5821 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     5881 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5941 agatcggcaa gtgcacaaac aatacttaaa taaatactac tcagtaataa cctatttctt
     6001 agcatttttg acgaaatttg ctattttgtt agagtctttt acaccatttg tctccacacc
     6061 tccgcttaca tcaacaccaa taacgccatt taatctaagc gcatcaccaa cattttctgg
     6121 cgtcagtcca ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca
     6181 gtcagaaatc gagttccaat ccaaaagttc acctgtccca cctgcttctg aatcaaacaa
     6241 gggaataaac gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc agtcttttgg
     6301 aaatacgagt cttttaataa ctggcaaacc gaggaactct tggtattctt gccacgactc
     6361 atctccatgc agttggacga tatcaatgcc gtaatcattg accagagcca aaacatcctc
     6421 cttaggttga ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac tatttttata
     6481 tgcttttaca agacttgaaa ttttccttgc aataaccggg tcaattgttc tctttctatt
     6541 gggcacacat ataataccca gcaagtcagc atcggaatct agagcacatt ctgcggcctc
     6601 tgtgctctgc aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga aattaataac
     6661 agacatactc caagctgcct ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca
     6721 ccaatgccct ccctcttggc cctctccttt tcttttttcg accgaattaa ttcttaatcg
     6781 gcaaaaaaag aaaagctccg gatcaagatt gtacgtaagg tgacaagcta tttttcaata
     6841 aagaatatct tccactactg ccatctggcg tcataactgc aaagtacaca tatattacga
     6901 tgctgttcta ttaaatgctt cctatattat atatatagta atgtcgtgat ctatggtgca
     6961 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     7021 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     7081 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
