LOCUS       sequence-341951-        8529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VP16-CIB1 in S. cerevisiae. Part of optogenetic system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8529)
  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 8529)
  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: pJL1
FEATURES             Location/Qualifiers
     source          1..8529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..683
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     protein_bind    2625..2742
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     protein_bind    2963..2979
                     /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     3037..3060
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3094..3159
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3166..3186
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5076..5095)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5077..5095)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5102..5119)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5102..5118)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5111..5133)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5259..5714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5346..5365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5556..5577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6070..6089)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6189..6211
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6249..6267)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6352..6456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6457..6525,6526..7317)
                     /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(6675..6694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7488..8076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7977..7996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8230..8247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8364..8385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8400..8405,8406..8423,8424..8430)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8438..8454
                     /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     8462..8478
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8462..8478
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8499..8517
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa cgattacatc aaaatccaca
      121 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat
      181 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta
      241 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt
      301 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac
      361 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca
      421 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga
      481 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
      541 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
      601 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
      661 tatttcaagg atataccatt ctaatgtctg cccctaagtt accaatagat agcggtttaa
      721 tataaggttt caacaagaaa tattttgtga aaatcaaaag ttaatagttt cactgtcgaa
      781 aatatgaaag tttaagtttt cagttcatcc cattgtagtt atcaaatttc tgtgtttccc
      841 gtatatgtgt gctctgtatt catcgctatc ttgaaactgg ttatgatcgt tgcgcatgtg
      901 acctgtatcg ctcaaaagag cccggcaccg gtaccgggca aaagccccta gcaagctaaa
      961 atgattcaac tttcgtagta atttgttcaa ccggtcacag taaagtcaat gactaacttt
     1021 tgaatcgagg atgtatttcc aattgaaagc aatgtatata taaaccattg tgttgaagct
     1081 tgtcttacag ttttcttgta tcctcccatt gttcatatag tacttaaagt accataatcc
     1141 ttaaagaaca aacttgaaag agaatacact atacacaact aataaataaa gaataatcat
     1201 ggctgtgacc aagacaattg tagttctacc aggtgaccat gttggtcaag aaatcactga
     1261 agaggccatt aaagttttaa atgctattca ggaatgtcgt ccagacaagg tcaatttcaa
     1321 gtttgatcat catttgatcg gtggtgctgc aattgatgcc actggtgttc cattaccaga
     1381 cgaagctttg gaggcctcta agaaagctga tgccgtgctt ctgggtgctg ttggtggtcc
     1441 aaaatggggt actggcgctg tcagaccaga acaaggtctt ttgaaaatcc gtaaggagtt
     1501 gcaattgtat gctaatctaa gaccatgtaa ttttgcatct gattccttac tagatctatc
     1561 gccattgaag cctgaaattg caagaggtac agatttcgtc gttgttagag aactagtggg
     1621 tggtatttat ttcggggaga gaaaagaaga tgaaggtgat ggtgtcgcct gggatagcga
     1681 aaagtattct gtgcccgaag ttcaaagaat cactagaatg gctgctttca tggctctgca
     1741 acacaaccca cctttaccta tttggtctct agataaagcc aacgttctag catcatcaag
     1801 attatggaga aaaacagtgg aagagacgat taagaatgaa ttcccacaat tgactgttca
     1861 acatcaattg attgattccg ctgctatgat cttggttaag aacccaactc atctaaatgg
     1921 tattattatc acaaacaata tgtttggtga tattatctcc gatgaagcct ccgtgatccc
     1981 aggttcgtta ggattacttc catctgcctc cttggcatct cttccagata agaatacggc
     2041 atttggttta tatgagccat gtcatggctc tgctccagac ttaccaaagg gaaaggttaa
     2101 ccccgtagca acaatcctat ctgctgctat gatgttgaag ttgtctctgg acctgttcga
     2161 agaaggtgaa attatagaac aagcagtcaa gaaagtgttg gattctggta tcagaacagc
     2221 ggatctaaag ggtaccaact ctactactga ggtcggtgat gcagtagcga aagctgtcag
     2281 ggaactatta gcttagactt agtagatgta cagttgcata cataagtctg aacagaaaga
     2341 agtaaatatc atatctagtt cggtttagaa atcagtgact atattggttt tgaccagatc
     2401 gcaatccttg caagcattta gtaatgacta tcaaaccggt gaatcatctg ataaggctat
     2461 attagcttgt gcattcgcat gtatcgggaa acgaacggta ccgggcccga gcagatccgc
     2521 caggcgtgta tatatagcgt ggatggccag gcaactttag tgctgacaca tacaggcata
     2581 tatatatgtg tgcgacgaca catgatcata tgggtaccga agtacggatt agaagccgcc
     2641 gagcgggtga cagccctccg aaggaagact ctcctccgtg cgtcctcgtc ttcaccggtc
     2701 gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca
     2761 atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt
     2821 caaatgaacg aatcaaatta acaaccatag gatgataatg cgattacttt tttagcctta
     2881 tttctggggt actgcagcag cgaagcgatg atttttgatc tattaacaga tatataaatg
     2941 caaaaactgt tgttgtgtgg aattgtgagc ggataacaat ttcacacaat attacttctt
     3001 attcaaatgt aataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc
     3061 aaggagaaaa accccttggg ccctaaaaaa atggactaca aagaccatga cggtgattat
     3121 aaagatcatg acatcgatta caaggatgac gatgacaaga tggcccccaa gaaaaagagg
     3181 aaggtgggca ttcacggggt gccgggtgga ctcgagggtt ctggttcctg cagggctcca
     3241 ccaactgatg tttcattggg tgatgaattg catttggatg gagaagatgt tgctatggct
     3301 catgctgatg cattagatga tttcgattta gacatgttgg gagatggtga ttctccaggt
     3361 ccaggtttca ctccacatga ttctgctcct tacggtgcat tggatatggc tgattttgaa
     3421 ttcgaacaaa tgttcactga tgctttagga attgatgaat atggtggtgg tgcggccgct
     3481 ggaggttctg gcggtggaag tggtggcgga ggtagcggtg gtggcagcgg tcccggggag
     3541 cgccccttcc agtgtcgcat ttgcatgcgg aacttttcga gacaatctaa tttgtcaaga
     3601 catacccgta ctcataccgg tgaaaaaccg tttcagtgtc ggatctgtat gcgaaatttc
     3661 tccagaaatg aacatttggt tttgcatcta cgtacgcaca ccggcgagaa gccattccaa
     3721 tgccgaatat gcatgcgcaa cttcagtcaa aaaactggtt tgagagttca cctaaaaacc
     3781 cacctgagag gatcctaagc gccaccgcgg tggagctcta agcaaatagc taaattatat
     3841 acgaattaat attatgatta agtgtttacg tgagtgcgat atttttatta ctatcttata
     3901 cagttgtata tactctataa aatgagttgt ctattaatta acgcgatgaa tgctttctgg
     3961 gtttacctct ccaacaactc tagtttactt ctcaatacat tcaattgtat ttgatttgtc
     4021 aatacttcat cattaatcaa ttctatagtt ttgtttttct cgtttatttc caaatttaat
     4081 gcatcaattt tattattcaa tttgtcgttg attttggtta atgattttat ggtttgatct
     4141 ctggcattga ttgtttgtgt tagtttttca ttattgataa ttaaattatt taagttagtt
     4201 atcaactcgg tgttttcaag tttcaagttt tcaatttctt tagagtttat tagatttgtc
     4261 aaagtttctg aattgcttga ttggtcctgt agaagagtat ttgttgttgt ggataattga
     4321 ttcaattttt gagacaattg ctggaaggcg ttgaaatatc tagcatcaat ctcatggttt
     4381 ttttcccgag agtctcgtag attcaattgt tttaatatat cttgggacca ctcttgattt
     4441 gaactcatgg aaattaaact gggtgttgtg ttgtggtgta atgattgtac cccctttgct
     4501 tataattgtg tggccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt
     4561 gcttaaaaag attctctttt tttatgatat ttgtacataa actttataaa tgaaattcat
     4621 aatagaaacg acacgaaatt acaaaatgga atatgttcat agggtagacg aaactatata
     4681 cgcaatctac atacatttat caagaaggag aaaaaggagg atgtaaagga atacaggtaa
     4741 gcaaattgat actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata
     4801 ttgacaagga ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg
     4861 attcctaatt tatatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa
     4921 acactcaatg acctgaccat ttgatggagt ttaagtcaat acctccttga accatttccc
     4981 ataatggtga aagttccctc aagaatttta ctctgtcaga aacggcctta acgacgtagg
     5041 ccggccattt aaatgtttaa acggccacgc tggcccccta tagtgagtcg tattacaatt
     5101 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     5161 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     5221 gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt
     5281 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5341 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5401 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5461 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5521 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5581 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5641 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5701 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5761 cgcatagggt aataactgat ataattaaat tgaagctcta atttgtgagt ttagtattag
     5821 gatctctacc cttggcgaaa agtcctctgc caacaatgat gatatctgat ccaccactta
     5881 caacttcgtc gacggttctg tactgctgac ccaatatgcg tatatatacc aatctaagtc
     5941 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag
     6001 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg
     6061 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     6121 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6181 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6241 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6301 ttagacgtcg gcgcgccgag gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6361 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6421 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6481 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6541 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6601 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6661 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6721 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6781 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6841 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6901 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6961 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7021 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7081 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7141 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7201 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7261 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7321 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7381 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7441 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7501 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7561 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7621 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7681 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7741 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7801 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7861 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7921 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7981 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8041 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8101 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8161 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     8221 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     8281 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     8341 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     8401 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     8461 acaggaaaca gctatgacca tgattacgcc aagctcggaa ttaaccctca ctaaagggaa
     8521 caaaagctg
//
