LOCUS       sequence-341751-        7626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 97-3 zinc finger array fused to VP16 in S. cerevisiae. 
            Parent plasmid for chromatin regulator library..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  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 7626)
  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: pJL50
FEATURES             Location/Qualifiers
     source          1..7626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             26..91
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             98..118
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1830..1849)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1831..1849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1856..1873)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1856..1872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1865..1887)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2013..2468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2100..2119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2310..2331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2824..2843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2943..2965
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3003..3021)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3106..3210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3211..3279,3280..4071)
                     /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(3429..3448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4242..4830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4731..4750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4984..5001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5118..5139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5154..5159,5160..5177,5178..5184)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5192..5208
                     /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     5197..5219
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5216..5232
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5216..5232
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5253..5271
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7223..7621
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 attacaaaag ggccctaaaa aaatggacta caaagaccat gacggtgatt ataaagatca
       61 tgacatcgat tacaaggatg acgatgacaa gatggccccc aagaaaaaga ggaaggtggg
      121 cattcacggg gtgccgggtg gaggttctgg ttctagaccc ggggagcgcc ccttccagtg
      181 tcgcatttgc atgcggaact tttcgagaca atctaatttg tcaagacata cccgtactca
      241 taccggtgaa aaaccgtttc agtgtcggat ctgtatgcga aatttctcca gaaatgaaca
      301 tttggttttg catctacgta cgcacaccgg cgagaagcca ttccaatgcc gaatatgcat
      361 gcgcaacttc agtcaaaaaa ctggtttgag agttcaccta aaaacccacc tgagaggatc
      421 cggaggttct ggcggtggaa gtggtggcgg aggtagcggt ggtggcagcg gttcctgcag
      481 ggctccacca actgatgttt cattgggtga tgaattgcat ttggatggag aagatgttgc
      541 tatggctcat gctgatgcat tagatgattt cgatttagac atgttgggag atggtgattc
      601 tccaggtcca ggtttcactc cacatgattc tgctccttac ggtgcattgg atatggctga
      661 ttttgaattc gaacaaatgt tcactgatgc tttaggaatt gatgaatatg gtggtggtta
      721 agcggccgcc accgcggtgg agctctaagc aaatagctaa attatatacg aattaatatt
      781 atgattaagt gtttacgtga gtgcgatatt tttattacta tcttatacag ttgtatatac
      841 tctataaaat gagttgtcta ttaattaacg cgatgaatgc tttctgggtt tacctctcca
      901 acaactctag tttacttctc aatacattca attgtatttg atttgtcaat acttcatcat
      961 taatcaattc tatagttttg tttttctcgt ttatttccaa atttaatgca tcaattttat
     1021 tattcaattt gtcgttgatt ttggttaatg attttatggt ttgatctctg gcattgattg
     1081 tttgtgttag tttttcatta ttgataatta aattatttaa gttagttatc aactcggtgt
     1141 tttcaagttt caagttttca atttctttag agtttattag atttgtcaaa gtttctgaat
     1201 tgcttgattg gtcctgtaga agagtatttg ttgttgtgga taattgattc aatttttgag
     1261 acaattgctg gaaggcgttg aaatatctag catcaatctc atggtttttt tcccgagagt
     1321 ctcgtagatt caattgtttt aatatatctt gggaccactc ttgatttgaa ctcatggaaa
     1381 ttaaactggg tgttgtgttg tggtgtaatg attgtacccc ctttgcttat aattgtgtgg
     1441 ttccgatgct gacttactgg gtattatatg tgtgcccaat agaaagagaa caattgaccc
     1501 ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt caggcactcc
     1561 gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg ctctggtcaa
     1621 tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcgag aataccaaga
     1681 gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca acatactact
     1741 cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag aaggccggcc
     1801 atttaaatgt ttaaacggcc acgctggccc cctatagtga gtcgtattac aattcactgg
     1861 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     1921 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     1981 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     2041 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2101 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2161 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2221 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2281 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2341 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2401 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     2461 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2521 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta ttaggatctc
     2581 tacccttggc gaaaagtcct ctgccaacaa tgatgatatc tgatccacca cttacaactt
     2641 cgtcgacggt tctgtactgc tgacccaata tgcgtatata taccaatcta agtctgtgct
     2701 ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc
     2761 gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct
     2821 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2881 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2941 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3001 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3061 gtcggcgcgc cgaggacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3181 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3241 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3301 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3361 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3421 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3481 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3541 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3601 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3661 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3721 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3781 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3841 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3901 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3961 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4021 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4081 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4141 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5101 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5161 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5221 aacagctatg accatgatta cgccaagctc ggaattaacc ctcactaaag ggaacaaaag
     5281 ctgggccggc catttaaatg tttaaacggc cacgctggcc atatttctac caatctctca
     5341 acactgagta atggtagtta taagaaagag accgagttag ggacagttag aggcggtgga
     5401 gatattcctt atggcatgtc tggcgatgat aaaacttttc aaacggcagc cccgatctaa
     5461 aagagctgac agggaaatgg tcagaaaaag aaacgtgcac ccgcccgtct ggacgcgccg
     5521 ctcacccgca cggcagagac caatcagtaa aaatcaacgg ttaacgacat tactatatat
     5581 ataatatagg aagcatttaa tagaacagca tcgtaatata tgtgtacttt gcagttatga
     5641 cgccagatgg cagtagtgga agatattctt tattgaaaaa tagcttgtca ccttacgtac
     5701 aatcttgatc cggagcccgt cgctttgaga gaaatcagaa gattccaaaa gtctactgaa
     5761 cttttgatca gaaagttgcc tttccaaaga ctagtcagag aaatcgccca agatttcaag
     5821 accgatctaa gattccaatc ctctgccatc ggtgctctac aggaatccgt tgaagcttac
     5881 ttggtctctt tgttcgaaga caccaacttg gccgctatcc acgctaagcg tgttaccatc
     5941 caaaagaagg atatcaagtt ggctagaaga ttgagaggtg aaagatccta agcacacagg
     6001 gattgttgta caattacttt ttatgatttt agatacacac aaatcttttc tacgattacc
     6061 tttcatttat tttcgtttca tgttttatta atcattcttt ttgtatattt cacatcgggt
     6121 ggggttttta tattaaattt taataacata atcttttaaa gtttctactt tctatcctgt
     6181 atattatttc catatctata tctatatttt caagcttgtt cacatcctta tcggaattgt
     6241 tccagcgttt ttgtacagaa aatgatgtta tggtaacgag ctatatcaga cctactagtc
     6301 ttatagagct gtacagagtc ttggaaacat aaccaacaca ccaacaagat gtcatttgat
     6361 tcgttactcg acaagaatga taagctggta aaagtttgcg ggattcaaac cgtcgaggct
     6421 gccgaaactg cgcttcaagc aggcgctgat ttgataggga tcatatgtgt ccccaacagg
     6481 aagcggacta tcgagagcgc tgtggctcgt gaaatatcca aattgattca caaatcaggt
     6541 actacaaagc tggtgggggt gttcaggaat caatctgttg aggacgtaca tcggctttct
     6601 gaggaatatg accttgacat aatccaatta catggtgatg aatcatggcc agagtactat
     6661 aacgtcatta agaaaccaat aatcaaaaga gtcatattcc ctagagatgt cgatgttgta
     6721 acacaagtgt gtcaaagaaa acccttggta tgtctaccat tgttcgactc cgaggcaggc
     6781 ggtacaggtg aaaagctgga ctggtccagt atttcctcct gggcaagcga acagaataac
     6841 gtattttata tactagcggg tggactaact gcagaaaacg tcatggaggc ggtggggttg
     6901 cctggtgtaa ttggtgttga cgtctcaggt ggtgtagaga ctgacggcgt caaagacaat
     6961 gacaagataa ttaaatatgt acaaaatgca aagaaacaat gaattacgaa gagaagtctc
     7021 aatgaggttt tacaaacaat cctataatac atatatgcca cttagatata ctttgaatac
     7081 tgaatatact gactcattat attctttaat tgtagtttct caaaaatatt catacaggtt
     7141 agctatgaat tcatgctata atttccattt tgttcatgta agatacacat agggataaaa
     7201 tgtgataact aatcagcggt accatagctt caaaatgttt ctactccttt tttactcttc
     7261 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     7321 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     7381 aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     7441 tcacgtttct ttttcttgaa aatttttttt ttgatttttt tctctttcga tgacctccca
     7501 ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt
     7561 tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa tctaatctaa
     7621 gtttta
//
