LOCUS       sequence-306294-        8392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrating plasmid to express MCP-ABI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8392)
  AUTHORS   Cunningham-Bryant D, Sun J, Fernandez B, Zalatan JG
  TITLE     CRISPR-Cas-Mediated Chemical Control of Transcriptional Dynamics in 
            Yeast.
  JOURNAL   Chembiochem. 2019 Jun 14;20(12):1519-1523. doi: 
            10.1002/cbic.201800823. Epub 2019 Apr 26.
  PUBMED    30710419
REFERENCE   2  (bases 1 to 8392)
  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"; doi: 
            "10.1002/cbic.201800823"; journalName: "Chembiochem"; date: 
            "2019-06-14- 14"; volume: "20"; issue: "12"; pages: "1519-1523"
COMMENT     Sequence Label: pJS033
FEATURES             Location/Qualifiers
     source          1..8392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        424..837
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             865..885
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3441..3460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3442..3460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3467..3484)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3467..3483)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3476..3498)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3624..4079
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3711..3730)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3921..3942
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4435..4454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4554..4576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4614..4632)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4822..4890,4891..5682)
                     /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(5040..5059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5853..6441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6342..6361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6595..6612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6729..6750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6765..6770,6771..6788,6789..6795)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6803..6819
                     /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     6808..6830
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6827..6843
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6827..6843
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6864..6882
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7208..7395
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 aaattaagta tataaagcag ttagtttctt ctgcttcttg tggttctggg tattcttgta
       61 atatttggaa taatagaaaa ccaagactac tgcctttctt ttcatattat agaggaagaa
      121 atacgcacga acacgatata gaggtaaatt ttttgcaaaa agataccaaa ctataaaaga
      181 aatagaagac taaactacaa aaaataaaga aagtaataca tatatccaat atgactgagt
      241 tagcagagca agtttccatg cctgaagggc aacaaagacg tggtggttac gttccacctc
      301 acatccgtaa cggtggtggc agaagaggcg attctgctcc tcgtggtggt ttcaacaacg
      361 gtggcggccg tggtggtttc ttcagaaaca acggtgaccg tggtggattc ggctttgggt
      421 accgggcccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
      481 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      541 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      601 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      661 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      721 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      781 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      841 taattacaaa tgtctctcga gatgcccaaa aagaaaagaa aagtgggtag tatggcttct
      901 aactttactc agttcgttct cgtcgacaat ggcggaactg gcgacgtgac tgtcgcccca
      961 agcaacttcg ctaacgggat cgctgaatgg atcagctcta actcgcgttc acaggcttac
     1021 aaagtaacct gtagcgttcg tcagagctct gcgcagaatc gcaaatacac catcaaagtc
     1081 gaggtgccta aaggcgcctg gcgttcgtac ttaaatatgg aactaaccat tccaattttc
     1141 gccacgaatt ccgactgcga gcttattgtt aaggcaatgc aaggtctcct aaaagatgga
     1201 aacccgattc cctcagcaat cgcagcaaac tccggcatct acggctccac gcgtgtgcct
     1261 ttgtatggtt ttacttcgat ttgtggaaga agacctgaga tggaagctgc tgtttcgact
     1321 ataccaagat tccttcaatc ttcctctggt tcgatgttag atggtcggtt tgatcctcaa
     1381 tccgccgctc atttcttcgg tgtttacgac ggccatggcg gttctcaggt agcgaactat
     1441 tgtagagaga ggatgcattt ggctttggcg gaggagatag ctaaggagaa accgatgctc
     1501 tgcgatggtg atacgtggct ggagaagtgg aagaaagctc ttttcaactc gttcctgaga
     1561 gttgactcgg agattgagtc agttgcgccg gagacggttg ggtcaacgtc ggtggttgcc
     1621 gttgttttcc cgtctcacat cttcgtcgct aactgcggtg actctagagc cgttctttgc
     1681 cgcggcaaaa ctgcacttcc attatccgtt gaccataaac cggatagaga agatgaagct
     1741 gcgaggattg aagccgcagg agggaaagtg attcagtgga atggagctcg tgttttcggt
     1801 gttctcgcca tgtcgagatc cattggcgat agatacttga aaccatccat cattcctgat
     1861 ccggaagtga cggctgtgaa gagagtaaaa gaagatgatt gtctgatttt ggcgagtgac
     1921 ggggtttggg atgtaatgac ggatgaagaa gcgtgtgaga tggcaaggaa gcggattctc
     1981 ttgtggcaca agaaaaacgc ggtggctggg gatgcatcgt tgctcgcgga tgagcggaga
     2041 aaggaaggga aagatcctgc ggcgatgtcc gcggctgagt atttgtcaaa gctggcgata
     2101 cagagaggaa gcaaagacaa cataagtgtg gtggtggttg atttgaaggc ggccgccacc
     2161 gcggtggagc tctaagcaaa tagctaaatt atatacgaat taatattatg attaagtgtt
     2221 tacgtgagtg cgatattttt attactatct tatacagttg tatatactct ataaaatgag
     2281 ttgtctatta attaacgcga tgaatgcttt ctgggtttac ctctccaaca actctagttt
     2341 acttctcaat acattcaatt gtatttgatt tgtcaatact tcatcattaa tcaattctat
     2401 agttttgttt ttctcgttta tttccaaatt taatgcatca attttattat tcaatttgtc
     2461 gttgattttg gttaatgatt ttatggtttg atctctggca ttgattgttt gtgttagttt
     2521 ttcattattg ataattaaat tatttaagtt agttatcaac tcggtgtttt caagtttcaa
     2581 gttttcaatt tctttagagt ttattagatt tgtcaaagtt tctgaattgc ttgattggtc
     2641 ctgtagaaga gtatttgttg ttgtggataa ttgattcaat ttttgagaca attgctggaa
     2701 ggcgttgaaa tatctagcat caatctcatg gtttttttcc cgagagtctc gtagattcaa
     2761 ttgttttaat atatcttggg accactcttg atttgaactc atggaaatta aactgggtgt
     2821 tgtgttgtgg tgtaatgatt gtaccccctt tgcttataat tgtgtggtga caccgattat
     2881 ttaaagctgc agcatacgat atatatacat gtgtatatat gtatacctat gaatgtcagt
     2941 aagtatgtat acgaacagta tgatactgaa gatgacaagg taatgcatca ttctatacgt
     3001 gtcattctga acgaggcgcg ctttcctttt ttctttttgc tttttctttt tttttctctt
     3061 gaactccaga aaaaaaatat aaaagagatg gaggaacggg aaaaagttag ttgtggtgat
     3121 aggtggcaag tggtattccg taagaacaac aagaaaagca tttcatatta tggctgaact
     3181 gagcgaacaa gtgcaaaatt taagcatcaa cgacaacaac gagaatggtt atgttcctcc
     3241 tcacttaaga ggaaaaccaa gaagtgccag aaataacagt agcaactaca ataacaacaa
     3301 cggcggctac aacggtggcc gtggcggtgg cagcttcttt agcaacaacc gtcgtggtgg
     3361 ttacggcaac ggtggtttct tcggtggaaa caacggtggc agcggccggc catttaaatg
     3421 tttaaacggc cacgctggcc ccctatagtg agtcgtatta caattcactg gccgtcgttt
     3481 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3541 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3601 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3661 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3721 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3781 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3841 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3901 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3961 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4021 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     4081 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agggtaataa
     4141 ctgatataat taaattgaag ctctaatttg tgagtttagt attaggatct ctacccttgg
     4201 cgaaaagtcc tctgccaaca atgatgatat ctgatccacc acttacaact tcgtcgacgg
     4261 ttctgtactg ctgacccaat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     4321 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa
     4381 aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat
     4441 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4501 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4561 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4621 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcggcgcg
     4681 ccgaggacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4861 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4921 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4981 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5041 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5101 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5161 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5221 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5281 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5341 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5401 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5461 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5521 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5581 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5641 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5701 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5761 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5821 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5881 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5941 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6001 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6061 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6481 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6541 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6601 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6661 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6721 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6781 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6841 gaccatgatt acgccaagct cggaattaac cctcactaaa gggaacaaaa gctgggccgg
     6901 ccatttaaat gtttaaacgg ccacgctggc cgctgcacgg tcctgttccc tagcatgtac
     6961 gtgagcgtat ttccttttaa accacgacgc tttgtcttca ttcaacgttt cccattgttt
     7021 ttttctacta ttgctttgct gtgggaaaaa cttatcgaaa gatgacgact ttttcttaat
     7081 tctcgtttta agagcttggt gagcgctagg agtcactgcc aggtatcgtt tgaacacggc
     7141 attagtcagg gaagtcataa cacagtcctt tcccgcaatt ttctttttct attactcttg
     7201 gcctcctcta gtacactcta tattttttta tgcctcggta atgattttca tttttttttt
     7261 tccacctagc ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta
     7321 tacattatat aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga
     7381 taaacgaagg caaagaggct ctatatgtat ataaagctgc ggatgccttc tgatttaatc
     7441 tagttctagt ttacttcagg tttacttaaa gttgagataa ttgaagcaaa gagaagcatc
     7501 gagcaaagga tggcgtttgt taagagggtt acgcaggaga cgaatataca gctggcgctg
     7561 gatcttgacg gtgggtctgt ttctgtacgg gagagcatac tgggcaagga atatgctagt
     7621 ggtgatgggc agaccatcca tgtgcacact ggagttgggt ttttggacca catgttgact
     7681 gcgctggcga agcatggcgg gtggtctctg atcctggagt gtatagggga cttgcacatc
     7741 gatgaccacc acactgttga ggactgtggg attgcgctgg gccaagcgtt caaggaggcg
     7801 cttggctccg tgcgtggtat caagaggttc gggcatgggt ttgcaccact ggacgaggcg
     7861 ctgagccgcg ctgtggttga cttctccaat aggcctttcg ccgtggtgga gctgggcctg
     7921 aagcgagaac gcataggcca gctatccaca gagatgatcc cgcacttctt ggagagtttc
     7981 gccactgagg cgcgtatcac catgcatgtg gactgtctgc ggggcaccaa cgaccaccac
     8041 cgctccgaat cagctttcaa ggcgctcgcc atcgccatca gagaggcaag aacacctacg
     8101 ggtcgcgatg acgttccatc cactaagggt gtcctagcat agaacacagc ccacagctac
     8161 caccatcaac aatatttata tagataacgt acacatagaa atcacacaaa cagagtattt
     8221 attcttaact acatgaacta ccatcagacc gtctgggcca ctatataatg tgctattcat
     8281 aaacgtgatc actctacgta gcaggcgacc ccaggtgaaa atttttcagc gagctgccag
     8341 attgtcaggt gaaaaactga aaaaaacttc tgggcgatga gcttgtggcg gg
//
