LOCUS       sequence-102114-        8762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCP-VP64 fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8762)
  AUTHORS   Zalatan JG, Lee ME, Almeida R, Gilbert LA, Whitehead EH, La Russa M,
            Tsai JC, Weissman JS, Dueber JE, Qi LS, Lim WA
  TITLE     Engineering Complex Synthetic Transcriptional Programs with CRISPR 
            RNA Scaffolds.
  JOURNAL   Cell. 2014 Dec 18. pii: S0092-8674(14)01570-0. doi: 
            10.1016/j.cell.2014.11.052.
  PUBMED    25533786
REFERENCE   2  (bases 1 to 8762)
  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. 2014 
            Dec 18. pii: S0092-8674(14)01570-0. doi: 
            10.1016/j.cell.2014.11.052."
COMMENT     Sequence Label: pJZC504
FEATURES             Location/Qualifiers
     source          1..8762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        314..501
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        2711..3415
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3443..3463
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3851..4000
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(5309..5328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5310..5328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5335..5352)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5335..5351)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5344..5366)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5492..5947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5579..5598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5789..5810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6303..6322)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6422..6444
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6482..6500)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6585..6689
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6690..6758,6759..7550)
                     /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(6908..6927)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7721..8309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8210..8229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8463..8480
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8597..8618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8633..8638,8639..8656,8657..8663)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8671..8687
                     /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     8676..8698
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8695..8711
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8695..8711
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8732..8750
                     /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 ctttgggtac
     1921 cgggccctaa aacaagaaga gggttgacta catcacgatg agggggatcg aagaaatgat
     1981 ggtaaatgaa ataggaaatc aaggagcatg aaggcaaaag acaaatataa gggtcgaacg
     2041 aaaaataaag tgaaaagtgt tgatatgatg tatttggctt tgcggcgccg aaaaaacgag
     2101 tttacgcaat tgcacaatca tgctgactct gtggcggacc cgcgctcttg ccggcccggc
     2161 gataacgctg ggcgtgaggc tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt
     2221 ttaccctgcg ctaaggggcg agattggaga agcaataaga atgccggttg gggttgcgat
     2281 gatgacgacc acgacaactg gtgtcattat ttaagttgcc gaaagaacct gagtgcattt
     2341 gcaacatgag tatactagaa gaatgagcca agacttgcga gacgcgagtt tgccggtggt
     2401 gcgaacaata gagcgaccat gaccttgaag gtgagacgcg cataaccgct agagtacttt
     2461 gaagaggaaa cagcaatagg gttgctacca gtataaatag acaggtacat acaacactgg
     2521 aaatggttgt ctgtttgagt acgctttcaa ttcatttggg tgtgcacttt attatgttac
     2581 aatatggaag ggaactttac acttctccta tgcacatata ttaattaaag tccaatgcta
     2641 gtagagaagg ggggtaacac ccctccgcgc tcttttccga tttttttcta aaccgtggaa
     2701 tatttcggat atccttttgt tgtttccggg tgtacaatat ggacttcctc ttttctggca
     2761 accaaaccca tacatcggga ttcctataat accttcgttg gtctccctaa catgtaggtg
     2821 gcggagggga gatatacaat agaacagata ccagacagga cataatgggc taaacaagac
     2881 tacaccaatt acactgcctc attgatggtg gtacataacg aactaatact gtagccctag
     2941 acttgatagc catcatcata tcgaagtttc actacccttt ttccatttgc catctattga
     3001 agtaataata ggcgcatgca acttcttttc tttttttttc ttttctctct cccccgttgt
     3061 tgtctcacca tatccgcaat gacaaaaaaa tgatggaaga cactaaagga aaaaattaac
     3121 gacaaagaca gcaccaacag atgtcgttgt tccagagctg atgaggggta tctcgaagca
     3181 cacgaaactt tttccttcct tcattcacgc acactactct ctaatgagca acggtatacg
     3241 gccttccttc cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt
     3301 ataaatagac ctgcaattat taatcttttg tttcctcgtc attgttctcg ttccctttct
     3361 tccttgtttc tttttctgca caatatttca agctatacca agcatacaat caactatctc
     3421 atatacaatg tctctcgaga tgcccaaaaa gaaaagaaaa gtgggtagta tgtccaaaac
     3481 catcgttctt tcggtcggcg aggctactcg cactctgact gagatccagt ccaccgcaga
     3541 ccgtcagatc ttcgaagaga aggtcgggcc tctggtgggt cggctgcgcc tcacggcttc
     3601 gctccgtcaa aacggagcca agaccgcgta tcgcgtcaac ctaaaactgg atcaggcgga
     3661 cgtcgttgat tccggacttc cgaaagtgcg ctacactcag gtatggtcgc acgacgtgac
     3721 aatcgttgcg aatagcaccg aggcctcgcg caaatcgttg tacgatttga ccaagtccct
     3781 cgtcgcgacc tcgcaggtcg aagatcttgt cgtcaacctt gtgccgctgg gccgtggctc
     3841 cggacgagcc gatgctctcg acgattttga cctcgatatg ctgggatccg acgctctcga
     3901 tgacttcgac cttgacatgc tggggagtga cgctctggac gacttcgatc tcgacatgct
     3961 gggctccgac gccctggatg acttcgatct ggacatgctc atcaactgag cggccgccac
     4021 cgcggtggag ctctaagcaa atagctaaat tatatacgaa ttaatattat gattaagtgt
     4081 ttacgtgagt gcgatatttt tattactatc ttatacagtt gtatatactc tataaaatga
     4141 gttgtctatt aattaacgcg atgaatgctt tctgggttta cctctccaac aactctagtt
     4201 tacttctcaa tacattcaat tgtatttgat ttgtcaatac ttcatcatta atcaattcta
     4261 tagttttgtt tttctcgttt atttccaaat ttaatgcatc aattttatta ttcaatttgt
     4321 cgttgatttt ggttaatgat tttatggttt gatctctggc attgattgtt tgtgttagtt
     4381 tttcattatt gataattaaa ttatttaagt tagttatcaa ctcggtgttt tcaagtttca
     4441 agttttcaat ttctttagag tttattagat ttgtcaaagt ttctgaattg cttgattggt
     4501 cctgtagaag agtatttgtt gttgtggata attgattcaa tttttgagac aattgctgga
     4561 aggcgttgaa atatctagca tcaatctcat ggtttttttc ccgagagtct cgtagattca
     4621 attgttttaa tatatcttgg gaccactctt gatttgaact catggaaatt aaactgggtg
     4681 ttgtgttgtg gtgtaatgat tgtaccccct ttgcttataa ttgtgtggtg acaccgatta
     4741 tttaaagctg cagcatacga tatatataca tgtgtatata tgtataccta tgaatgtcag
     4801 taagtatgta tacgaacagt atgatactga agatgacaag gtaatgcatc attctatacg
     4861 tgtcattctg aacgaggcgc gctttccttt tttctttttg ctttttcttt ttttttctct
     4921 tgaactccag aaaaaaaata taaaagagat ggaggaacgg gaaaaagtta gttgtggtga
     4981 taggtggcaa gtggtattcc gtaagaacaa caagaaaagc atttcatatt atggctgaac
     5041 tgagcgaaca agtgcaaaat ttaagcatca acgacaacaa cgagaatggt tatgttcctc
     5101 ctcacttaag aggaaaacca agaagtgcca gaaataacag tagcaactac aataacaaca
     5161 acggcggcta caacggtggc cgtggcggtg gcagcttctt tagcaacaac cgtcgtggtg
     5221 gttacggcaa cggtggtttc ttcggtggaa acaacggtgg cagcggccgg ccatttaaat
     5281 gtttaaacgg ccacgctggc ccaattcgcc ctatagtgag tcgtattaca attcactggc
     5341 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5401 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5461 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     5521 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5581 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5641 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5701 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5761 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5821 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5941 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     6001 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat taggatctct
     6061 acccttggcg aaaagtcctc tgccaacaat gatgatatct gatccaccac ttacaacttc
     6121 gtcgacggtt ctgtactgct gacccaatat gcgtatatat accaatctaa gtctgtgctc
     6181 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg
     6241 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa aggtggtatg gtgcactctc
     6301 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6361 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6421 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     6481 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6541 tcggcgcgcc gaggacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6601 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6661 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6721 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6781 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6841 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6901 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6961 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7021 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7081 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     7141 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7201 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     7261 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7321 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7381 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7441 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7501 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7561 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7621 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7681 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7741 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7801 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7861 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7921 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7981 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8041 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     8101 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     8161 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     8221 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     8281 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8341 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     8401 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     8461 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     8521 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     8581 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     8641 ttatgcttcc ggctcctatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     8701 acagctatga ccatgattac gccaagctcg gaattaaccc tcactaaagg gaacaaaagc
     8761 tg
//
