LOCUS       sequence-357458-        8987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for CIB1 fused to VP16 in S. cerevisiae. Part of optogenetic
            system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8987)
  AUTHORS   Lee JB, Caywood LM, Lo JY, Levering N, Keung AJ
  TITLE     Mapping the dynamic transfer functions of eukaryotic gene 
            regulation.
  JOURNAL   Cell Syst. 2021 Aug 24. pii: S2405-4712(21)00291-X. doi: 
            10.1016/j.cels.2021.08.003.
  PUBMED    34469745
REFERENCE   2  (bases 1 to 8987)
  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: "Cell Syst. 
            2021 Aug 24. pii: S2405-4712(21)00291-X. doi: 
            10.1016/j.cels.2021.08.003."
COMMENT     Sequence Label: pJL32
FEATURES             Location/Qualifiers
     source          1..8987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        314..501
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     protein_bind    2264..2381
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     protein_bind    2602..2618
                     /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     2676..2699
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2733..2798
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2805..2825
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2898..3407
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCN
                     EKKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     primer_bind     complement(5534..5553)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5535..5553)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5560..5577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5560..5576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5569..5591)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5717..6172
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5804..5823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6014..6035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6528..6547)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6647..6669
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6707..6725)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6810..6914
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6915..6983,6984..7775)
                     /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(7133..7152)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7946..8534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8435..8454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8688..8705
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8822..8843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8858..8863,8864..8881,8882..8888)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8896..8912
                     /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     8920..8936
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8920..8936
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8957..8975
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgct gcacggtcct gttccctagc
       61 atgtacgtga gcgtatttcc ttttaaacca cgacgctttg tcttcattca acgtttccca
      121 ttgttttttt ctactattgc tttgctgtgg gaaaaactta tcgaaagatg acgacttttt
      181 cttaattctc gttttaagag cttggtgagc gctaggagtc actgccaggt atcgtttgaa
      241 cacggcatta gtcagggaag tcataacaca gtcctttccc gcaattttct ttttctatta
      301 ctcttggcct cctctagtac actctatatt tttttatgcc tcggtaatga ttttcatttt
      361 tttttttcca cctagcggat gactcttttt ttttcttagc gattggcatt atcacataat
      421 gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag
      481 gcaagataaa cgaaggcaaa gaggctctat atgtatataa agctgcggat gccttctgat
      541 ttaatctagt tctagtttac ttcaggttta cttaaagttg agataattga agcaaagaga
      601 agcatcgagc aaaggatggc gtttgttaag agggttacgc aggagacgaa tatacagctg
      661 gcgctggatc ttgacggtgg gtctgtttct gtacgggaga gcatactggg caaggaatat
      721 gctagtggtg atgggcagac catccatgtg cacactggag ttgggttttt ggaccacatg
      781 ttgactgcgc tggcgaagca tggcgggtgg tctctgatcc tggagtgtat aggggacttg
      841 cacatcgatg accaccacac tgttgaggac tgtgggattg cgctgggcca agcgttcaag
      901 gaggcgcttg gctccgtgcg tggtatcaag aggttcgggc atgggtttgc accactggac
      961 gaggcgctga gccgcgctgt ggttgacttc tccaataggc ctttcgccgt ggtggagctg
     1021 ggcctgaagc gagaacgcat aggccagcta tccacagaga tgatcccgca cttcttggag
     1081 agtttcgcca ctgaggcgcg tatcaccatg catgtggact gtctgcgggg caccaacgac
     1141 caccaccgct ccgaatcagc tttcaaggcg ctcgccatcg ccatcagaga ggcaagaaca
     1201 cctacgggtc gcgatgacgt tccatccact aagggtgtcc tagcatagaa cacagcccac
     1261 agctaccacc atcaacaata tttatataga taacgtacac atagaaatca cacaaacaga
     1321 gtatttattc ttaactacat gaactaccat cagaccgtct gggccactat ataatgtgct
     1381 attcataaac gtgatcactc tacgtagcag gcgaccccag gtgaaaattt ttcagcgagc
     1441 tgccagattg tcaggtgaaa aactgaaaaa aacttctggg cgatgagctt gtggcgggaa
     1501 attaagtata taaagcagtt agtttcttct gcttcttgtg gttctgggta ttcttgtaat
     1561 atttggaata atagaaaacc aagactactg cctttctttt catattatag aggaagaaat
     1621 acgcacgaac acgatataga ggtaaatttt ttgcaaaaag ataccaaact ataaaagaaa
     1681 tagaagacta aactacaaaa aataaagaaa gtaatacata tatccaatat gactgagtta
     1741 gcagagcaag tttccatgcc tgaagggcaa caaagacgtg gtggttacgt tccacctcac
     1801 atccgtaacg gtggtggcag aagaggcgat tctgctcctc gtggtggttt caacaacggt
     1861 ggcggccgtg gtggtttctt cagaaacaac ggtgaccgtg gtggattcgg ctttggcgaa
     1921 ttcggatcca ctagtactta gtagatgtac agttgcatac ataagtctga acagaaagaa
     1981 gtaaatatca tatctagttc ggtttagaaa tcagtgacta tattggtttt gaccagatcg
     2041 caatccttgc aagcatttag taatgactat caaaccggtg aatcatctga taaggctata
     2101 ttagcttgtg cattcgcatg tatcgggaaa cgaacggtac cgggcccgag cagatccgcc
     2161 aggcgtgtat atatagcgtg gatggccagg caactttagt gctgacacat acaggcatat
     2221 atatatgtgt gcgacgacac atgatcatat gggtaccgaa gtacggatta gaagccgccg
     2281 agcgggtgac agccctccga aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg
     2341 cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa
     2401 tactagcttt tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc
     2461 aaatgaacga atcaaattaa caaccatagg atgataatgc gattactttt ttagccttat
     2521 ttctggggta ctgcagcagc gaagcgatga tttttgatct attaacagat atataaatgc
     2581 aaaaactgtt gttgtgtgga attgtgagcg gataacaatt tcacacaata ttacttctta
     2641 ttcaaatgta ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca
     2701 aggagaaaaa ccccttgggc cctaaaaaaa tggactacaa agaccatgac ggtgattata
     2761 aagatcatga catcgattac aaggatgacg atgacaagat ggcccccaag aaaaagagga
     2821 aggtgggcat tcacggggtg ccgggtggac tcgagggttc tggttcatgt agggcgagcg
     2881 ccgaagctag cgccaccatg aatggagcta taggaggtga ccttttgctc aattttcctg
     2941 acatgtcggt cctagagcgc caaagggctc acctcaagta cctcaatccc acctttgatt
     3001 ctcctctcgc cggcttcttt gccgattctt caatgattac cggcggcgag atggacagct
     3061 atctttcgac tgccggtttg aatcttccga tgatgtacgg tgagacgacg gtggaaggtg
     3121 attcaagact ctcaatttcg ccggaaacga cgcttgggac tggaaatttc aagaaacgga
     3181 agtttgatac agagactaag gattgtaatg agaagaagaa gaagatgacg atgaacagag
     3241 atgacctagt agaagaagga gaagaagaga agtcgaaaat aacagagcaa aacaatggga
     3301 gcacaaaaag catcaagaag atgaaacaca aagccaagaa agaagagaac aatttctcta
     3361 atgattcatc taaagtgacg aaggaattgg agaaaacgga ttatattcat gttcgtgcac
     3421 gacgaggcca agccactgat agtcacagca tagcagaacg agttagaaga gaaaagatca
     3481 gtgagagaat gaagtttcta caagatttgg ttcctggatg cgacaagatc acaggcaaag
     3541 cagggatgct tgatgaaatc attaactatg ttcagtctct tcagagacaa atcgagttct
     3601 tatcgatgaa actagcaatt gtgaatccaa ggccggattt tgatatggat gacatttttg
     3661 ccaaagaggt tgcctcaact ccaatgactg tggtgccatc tcctgaaatg gttctttccg
     3721 gttattctca tgagatggtt cactctggtt attctagtga gatggttaac tccggttacc
     3781 ttcatgtcaa tccaatgcag caagtgaata ccagttctga tccattgtca tgcttcaaca
     3841 atggcgaagc tccttcgatg tgggactctc atgtgcagaa tctctatggc aatttaggag
     3901 taccggtcat cgagctcgag ctgcagatga atcgtagata cggaggttct ggcggtggaa
     3961 gtggtggcgg aggtagcggt ggtggcagcg gttcctgcag ggctccacca actgatgttt
     4021 cattgggtga tgaattgcat ttggatggag aagatgttgc tatggctcat gctgatgcat
     4081 tagatgattt cgatttagac atgttgggag atggtgattc tccaggtcca ggtttcactc
     4141 cacatgattc tgctccttac ggtgcattgg atatggctga ttttgaattc gaacaaatgt
     4201 tcactgatgc tttaggaatt gatgaatatg gtggtggtta agcggccgcc accgcggtgg
     4261 agctctaagc aaatagctaa attatatacg aattaatatt atgattaagt gtttacgtga
     4321 gtgcgatatt tttattacta tcttatacag ttgtatatac tctataaaat gagttgtcta
     4381 ttaattaacg cgatgaatgc tttctgggtt tacctctcca acaactctag tttacttctc
     4441 aatacattca attgtatttg atttgtcaat acttcatcat taatcaattc tatagttttg
     4501 tttttctcgt ttatttccaa atttaatgca tcaattttat tattcaattt gtcgttgatt
     4561 ttggttaatg attttatggt ttgatctctg gcattgattg tttgtgttag tttttcatta
     4621 ttgataatta aattatttaa gttagttatc aactcggtgt tttcaagttt caagttttca
     4681 atttctttag agtttattag atttgtcaaa gtttctgaat tgcttgattg gtcctgtaga
     4741 agagtatttg ttgttgtgga taattgattc aatttttgag acaattgctg gaaggcgttg
     4801 aaatatctag catcaatctc atggtttttt tcccgagagt ctcgtagatt caattgtttt
     4861 aatatatctt gggaccactc ttgatttgaa ctcatggaaa ttaaactggg tgttgtgttg
     4921 tggtgtaatg attgtacccc ctttgcttat aattgtgtgg tgacaccgat tatttaaagc
     4981 tgcagcatac gatatatata catgtgtata tatgtatacc tatgaatgtc agtaagtatg
     5041 tatacgaaca gtatgatact gaagatgaca aggtaatgca tcattctata cgtgtcattc
     5101 tgaacgaggc gcgctttcct tttttctttt tgctttttct ttttttttct cttgaactcc
     5161 agaaaaaaaa tataaaagag atggaggaac gggaaaaagt tagttgtggt gataggtggc
     5221 aagtggtatt ccgtaagaac aacaagaaaa gcatttcata ttatggctga actgagcgaa
     5281 caagtgcaaa atttaagcat caacgacaac aacgagaatg gttatgttcc tcctcactta
     5341 agaggaaaac caagaagtgc cagaaataac agtagcaact acaataacaa caacggcggc
     5401 tacaacggtg gccgtggcgg tggcagcttc tttagcaaca accgtcgtgg tggttacggc
     5461 aacggtggtt tcttcggtgg aaacaacggt ggcagcggcc ggccatttaa atgtttaaac
     5521 ggccacgctg gccccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     5581 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5641 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5701 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5761 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5821 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5881 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5941 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6001 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6061 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6121 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     6181 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat
     6241 aattaaattg aagctctaat ttgtgagttt agtattagga tctctaccct tggcgaaaag
     6301 tcctctgcca acaatgatga tatctgatcc accacttaca acttcgtcga cggttctgta
     6361 ctgctgaccc aatatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     6421 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga
     6481 ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct
     6541 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6601 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6661 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6721 ctatttttat aggttaatgt catgataata atggtttctt agacgtcggc gcgccgagga
     6781 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6841 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6901 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6961 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     7021 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7081 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7141 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7201 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     7261 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7321 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7381 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7441 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     7501 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7561 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7621 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7681 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7741 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7801 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7861 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7921 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7981 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8041 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     8101 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8161 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8221 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8281 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     8341 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     8401 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8461 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8521 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8581 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8641 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8701 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     8761 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     8821 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     8881 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     8941 attacgccaa gctcggaatt aaccctcact aaagggaaca aaagctg
//
