LOCUS       sequence-341771-        8750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CIB1 in S. cerevisiae. Includes inducible lac operon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8750)
  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 8750)
  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: pJL45
FEATURES             Location/Qualifiers
     source          1..8750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(674..693)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(675..693)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(700..717)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(700..716)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(709..731)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      857..1312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(944..963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1154..1175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1668..1687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1787..1809
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1847..1865)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1950..2054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2055..2123,2124..2915)
                     /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(2273..2292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3086..3674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3575..3594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3828..3845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3962..3983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3998..4003,4004..4021,4022..4028)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4036..4052
                     /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     4060..4076
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4060..4076
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4097..4115
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4441..4628
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     protein_bind    6391..6508
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     protein_bind    6729..6745
                     /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     6803..6826
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             6860..6925
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6932..6952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7025..7534
                     /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"
ORIGIN
        1 aatatatctt gggaccactc ttgatttgaa ctcatggaaa ttaaactggg tgttgtgttg
       61 tggtgtaatg attgtacccc ctttgcttat aattgtgtgg tgacaccgat tatttaaagc
      121 tgcagcatac gatatatata catgtgtata tatgtatacc tatgaatgtc agtaagtatg
      181 tatacgaaca gtatgatact gaagatgaca aggtaatgca tcattctata cgtgtcattc
      241 tgaacgaggc gcgctttcct tttttctttt tgctttttct ttttttttct cttgaactcc
      301 agaaaaaaaa tataaaagag atggaggaac gggaaaaagt tagttgtggt gataggtggc
      361 aagtggtatt ccgtaagaac aacaagaaaa gcatttcata ttatggctga actgagcgaa
      421 caagtgcaaa atttaagcat caacgacaac aacgagaatg gttatgttcc tcctcactta
      481 agaggaaaac caagaagtgc cagaaataac agtagcaact acaataacaa caacggcggc
      541 tacaacggtg gccgtggcgg tggcagcttc tttagcaaca accgtcgtgg tggttacggc
      601 aacggtggtt tcttcggtgg aaacaacggt ggcagcggcc ggccatttaa atgtttaaac
      661 ggccacgctg gccccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
      721 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      781 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      841 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
      901 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
      961 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1021 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1081 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1141 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1201 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1261 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     1321 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat
     1381 aattaaattg aagctctaat ttgtgagttt agtattagga tctctaccct tggcgaaaag
     1441 tcctctgcca acaatgatga tatctgatcc accacttaca acttcgtcga cggttctgta
     1501 ctgctgaccc aatatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     1561 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga
     1621 ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct
     1681 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1741 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1801 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     1861 ctatttttat aggttaatgt catgataata atggtttctt agacgtcggc gcgccgagga
     1921 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1981 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2041 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2101 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2161 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2221 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2281 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2341 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2401 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2461 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2521 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2581 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2641 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2701 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2761 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2821 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2881 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2941 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3001 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3061 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3241 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3841 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3901 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3961 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     4021 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     4081 attacgccaa gctcggaatt aaccctcact aaagggaaca aaagctgggc cggccattta
     4141 aatgtttaaa cggccacgct ggccgctgca cggtcctgtt ccctagcatg tacgtgagcg
     4201 tatttccttt taaaccacga cgctttgtct tcattcaacg tttcccattg tttttttcta
     4261 ctattgcttt gctgtgggaa aaacttatcg aaagatgacg actttttctt aattctcgtt
     4321 ttaagagctt ggtgagcgct aggagtcact gccaggtatc gtttgaacac ggcattagtc
     4381 agggaagtca taacacagtc ctttcccgca attttctttt tctattactc ttggcctcct
     4441 ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt ttttccacct
     4501 agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa ttatacatta
     4561 tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca agataaacga
     4621 aggcaaagag gctctatatg tatataaagc tgcggatgcc ttctgattta atctagttct
     4681 agtttacttc aggtttactt aaagttgaga taattgaagc aaagagaagc atcgagcaaa
     4741 ggatggcgtt tgttaagagg gttacgcagg agacgaatat acagctggcg ctggatcttg
     4801 acggtgggtc tgtttctgta cgggagagca tactgggcaa ggaatatgct agtggtgatg
     4861 ggcagaccat ccatgtgcac actggagttg ggtttttgga ccacatgttg actgcgctgg
     4921 cgaagcatgg cgggtggtct ctgatcctgg agtgtatagg ggacttgcac atcgatgacc
     4981 accacactgt tgaggactgt gggattgcgc tgggccaagc gttcaaggag gcgcttggct
     5041 ccgtgcgtgg tatcaagagg ttcgggcatg ggtttgcacc actggacgag gcgctgagcc
     5101 gcgctgtggt tgacttctcc aataggcctt tcgccgtggt ggagctgggc ctgaagcgag
     5161 aacgcatagg ccagctatcc acagagatga tcccgcactt cttggagagt ttcgccactg
     5221 aggcgcgtat caccatgcat gtggactgtc tgcggggcac caacgaccac caccgctccg
     5281 aatcagcttt caaggcgctc gccatcgcca tcagagaggc aagaacacct acgggtcgcg
     5341 atgacgttcc atccactaag ggtgtcctag catagaacac agcccacagc taccaccatc
     5401 aacaatattt atatagataa cgtacacata gaaatcacac aaacagagta tttattctta
     5461 actacatgaa ctaccatcag accgtctggg ccactatata atgtgctatt cataaacgtg
     5521 atcactctac gtagcaggcg accccaggtg aaaatttttc agcgagctgc cagattgtca
     5581 ggtgaaaaac tgaaaaaaac ttctgggcga tgagcttgtg gcgggaaatt aagtatataa
     5641 agcagttagt ttcttctgct tcttgtggtt ctgggtattc ttgtaatatt tggaataata
     5701 gaaaaccaag actactgcct ttcttttcat attatagagg aagaaatacg cacgaacacg
     5761 atatagaggt aaattttttg caaaaagata ccaaactata aaagaaatag aagactaaac
     5821 tacaaaaaat aaagaaagta atacatatat ccaatatgac tgagttagca gagcaagttt
     5881 ccatgcctga agggcaacaa agacgtggtg gttacgttcc acctcacatc cgtaacggtg
     5941 gtggcagaag aggcgattct gctcctcgtg gtggtttcaa caacggtggc ggccgtggtg
     6001 gtttcttcag aaacaacggt gaccgtggtg gattcggctt tggcgaattc ggatccacta
     6061 gtacttagta gatgtacagt tgcatacata agtctgaaca gaaagaagta aatatcatat
     6121 ctagttcggt ttagaaatca gtgactatat tggttttgac cagatcgcaa tccttgcaag
     6181 catttagtaa tgactatcaa accggtgaat catctgataa ggctatatta gcttgtgcat
     6241 tcgcatgtat cgggaaacga acggtaccgg gcccgagcag atccgccagg cgtgtatata
     6301 tagcgtggat ggccaggcaa ctttagtgct gacacataca ggcatatata tatgtgtgcg
     6361 acgacacatg atcatatggg taccgaagta cggattagaa gccgccgagc gggtgacagc
     6421 cctccgaagg aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt tcctgaaacg
     6481 cagatgtgcc tcgcgccgca ctgctccgaa caataaagat tctacaatac tagcttttat
     6541 ggttatgaag aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa tgaacgaatc
     6601 aaattaacaa ccataggatg ataatgcgat tactttttta gccttatttc tggggtactg
     6661 cagcagcgaa gcgatgattt ttgatctatt aacagatata taaatgcaaa aactgttgtt
     6721 gtgtggaatt gtgagcggat aacaatttca cacaatatta cttcttattc aaatgtaata
     6781 aaagtatcaa caaaaaattg ttaatatacc tctatacttt aacgtcaagg agaaaaaccc
     6841 cttgggccct aaaaaaatgg actacaaaga ccatgacggt gattataaag atcatgacat
     6901 cgattacaag gatgacgatg acaagatggc ccccaagaaa aagaggaagg tgggcattca
     6961 cggggtgccg ggtggactcg agggttctgg ttcatgtagg gcgagcgccg aagctagcgc
     7021 caccatgaat ggagctatag gaggtgacct tttgctcaat tttcctgaca tgtcggtcct
     7081 agagcgccaa agggctcacc tcaagtacct caatcccacc tttgattctc ctctcgccgg
     7141 cttctttgcc gattcttcaa tgattaccgg cggcgagatg gacagctatc tttcgactgc
     7201 cggtttgaat cttccgatga tgtacggtga gacgacggtg gaaggtgatt caagactctc
     7261 aatttcgccg gaaacgacgc ttgggactgg aaatttcaag aaacggaagt ttgatacaga
     7321 gactaaggat tgtaatgaga agaagaagaa gatgacgatg aacagagatg acctagtaga
     7381 agaaggagaa gaagagaagt cgaaaataac agagcaaaac aatgggagca caaaaagcat
     7441 caagaagatg aaacacaaag ccaagaaaga agagaacaat ttctctaatg attcatctaa
     7501 agtgacgaag gaattggaga aaacggatta tattcatgtt cgtgcacgac gaggccaagc
     7561 cactgatagt cacagcatag cagaacgagt tagaagagaa aagatcagtg agagaatgaa
     7621 gtttctacaa gatttggttc ctggatgcga caagatcaca ggcaaagcag ggatgcttga
     7681 tgaaatcatt aactatgttc agtctcttca gagacaaatc gagttcttat cgatgaaact
     7741 agcaattgtg aatccaaggc cggattttga tatggatgac atttttgcca aagaggttgc
     7801 ctcaactcca atgactgtgg tgccatctcc tgaaatggtt ctttccggtt attctcatga
     7861 gatggttcac tctggttatt ctagtgagat ggttaactcc ggttaccttc atgtcaatcc
     7921 aatgcagcaa gtgaatacca gttctgatcc attgtcatgc ttcaacaatg gcgaagctcc
     7981 ttcgatgtgg gactctcatg tgcagaatct ctatggcaat ttaggagtac cggtcatcga
     8041 gctcgagctg cagatgaatc gtagatacgg aggttctggc ggtggaagtg gtggcggagg
     8101 tagcggtggt ggcagcggtt cctgcaggta agcggccgcc accgcggtgg agctctaagc
     8161 aaatagctaa attatatacg aattaatatt atgattaagt gtttacgtga gtgcgatatt
     8221 tttattacta tcttatacag ttgtatatac tctataaaat gagttgtcta ttaattaacg
     8281 cgatgaatgc tttctgggtt tacctctcca acaactctag tttacttctc aatacattca
     8341 attgtatttg atttgtcaat acttcatcat taatcaattc tatagttttg tttttctcgt
     8401 ttatttccaa atttaatgca tcaattttat tattcaattt gtcgttgatt ttggttaatg
     8461 attttatggt ttgatctctg gcattgattg tttgtgttag tttttcatta ttgataatta
     8521 aattatttaa gttagttatc aactcggtgt tttcaagttt caagttttca atttctttag
     8581 agtttattag atttgtcaaa gtttctgaat tgcttgattg gtcctgtaga agagtatttg
     8641 ttgttgtgga taattgattc aatttttgag acaattgctg gaaggcgttg aaatatctag
     8701 catcaatctc atggtttttt tcccgagagt ctcgtagatt caattgtttt
//
