LOCUS       sequence-102113-        8747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MCP-VP64 fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8747)
  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 8747)
  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: pJZC522
FEATURES             Location/Qualifiers
     source          1..8747
                     /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             3836..3985
                     /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(5294..5313)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5295..5313)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5320..5337)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5320..5336)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5329..5351)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5477..5932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5564..5583)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5774..5795
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6288..6307)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6407..6429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6467..6485)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6570..6674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6675..6743,6744..7535)
                     /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(6893..6912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7706..8294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8195..8214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8448..8465
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8582..8603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8618..8623,8624..8641,8642..8648)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8656..8672
                     /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     8661..8683
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8680..8696
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8680..8696
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8717..8735
                     /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 tggcttctaa
     3481 ctttactcag ttcgttctcg tcgacaatgg cggaactggc gacgtgactg tcgccccaag
     3541 caacttcgct aacgggatcg ctgaatggat cagctctaac tcgcgttcac aggcttacaa
     3601 agtaacctgt agcgttcgtc agagctctgc gcagaatcgc aaatacacca tcaaagtcga
     3661 ggtgcctaaa ggcgcctggc gttcgtactt aaatatggaa ctaaccattc caattttcgc
     3721 cacgaattcc gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa
     3781 cccgattccc tcagcaatcg cagcaaactc cggcatctac ggctccggac gagccgatgc
     3841 tctcgacgat tttgacctcg atatgctggg atccgacgct ctcgatgact tcgaccttga
     3901 catgctgggg agtgacgctc tggacgactt cgatctcgac atgctgggct ccgacgccct
     3961 ggatgacttc gatctggaca tgctcatcaa ctgagcggcc gccaccgcgg tggagctcta
     4021 agcaaatagc taaattatat acgaattaat attatgatta agtgtttacg tgagtgcgat
     4081 atttttatta ctatcttata cagttgtata tactctataa aatgagttgt ctattaatta
     4141 acgcgatgaa tgctttctgg gtttacctct ccaacaactc tagtttactt ctcaatacat
     4201 tcaattgtat ttgatttgtc aatacttcat cattaatcaa ttctatagtt ttgtttttct
     4261 cgtttatttc caaatttaat gcatcaattt tattattcaa tttgtcgttg attttggtta
     4321 atgattttat ggtttgatct ctggcattga ttgtttgtgt tagtttttca ttattgataa
     4381 ttaaattatt taagttagtt atcaactcgg tgttttcaag tttcaagttt tcaatttctt
     4441 tagagtttat tagatttgtc aaagtttctg aattgcttga ttggtcctgt agaagagtat
     4501 ttgttgttgt ggataattga ttcaattttt gagacaattg ctggaaggcg ttgaaatatc
     4561 tagcatcaat ctcatggttt ttttcccgag agtctcgtag attcaattgt tttaatatat
     4621 cttgggacca ctcttgattt gaactcatgg aaattaaact gggtgttgtg ttgtggtgta
     4681 atgattgtac cccctttgct tataattgtg tggtgacacc gattatttaa agctgcagca
     4741 tacgatatat atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga
     4801 acagtatgat actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga
     4861 ggcgcgcttt ccttttttct ttttgctttt tctttttttt tctcttgaac tccagaaaaa
     4921 aaatataaaa gagatggagg aacgggaaaa agttagttgt ggtgataggt ggcaagtggt
     4981 attccgtaag aacaacaaga aaagcatttc atattatggc tgaactgagc gaacaagtgc
     5041 aaaatttaag catcaacgac aacaacgaga atggttatgt tcctcctcac ttaagaggaa
     5101 aaccaagaag tgccagaaat aacagtagca actacaataa caacaacggc ggctacaacg
     5161 gtggccgtgg cggtggcagc ttctttagca acaaccgtcg tggtggttac ggcaacggtg
     5221 gtttcttcgg tggaaacaac ggtggcagcg gccggccatt taaatgttta aacggccacg
     5281 ctggcccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     5341 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5401 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5461 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5521 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5581 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5641 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5701 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5761 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5821 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5881 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     5941 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat
     6001 aattaaattg aagctctaat ttgtgagttt agtattagga tctctaccct tggcgaaaag
     6061 tcctctgcca acaatgatga tatctgatcc accacttaca acttcgtcga cggttctgta
     6121 ctgctgaccc aatatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     6181 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga
     6241 ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct
     6301 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6361 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6421 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6481 ctatttttat aggttaatgt catgataata atggtttctt agacgtcggc gcgccgagga
     6541 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6601 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6661 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6721 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6781 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6841 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6901 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6961 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     7021 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7081 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7141 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7201 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     7261 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7321 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7381 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7441 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7501 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7561 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7621 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7681 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7741 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7801 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7861 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7921 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7981 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8041 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     8101 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     8161 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8221 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8281 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8341 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8401 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8461 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     8521 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     8581 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     8641 ctatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     8701 attacgccaa gctcggaatt aaccctcact aaagggaaca aaagctg
//
