LOCUS       sequence-305866-        8669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrating plasmid for pTEF1-mCherry reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8669)
  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 8669)
  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: pJZC533
FEATURES             Location/Qualifiers
     source          1..8669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1030..2386
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1030..1373
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1374..2183
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1441..1460)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2189..2386
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        2434..2847
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(5216..5235)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5217..5235)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5242..5259)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5242..5258)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5251..5273)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5399..5854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5486..5505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5696..5717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6210..6229)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6329..6351
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6389..6407)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6492..6596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6597..6665,6666..7457)
                     /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(6815..6834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7628..8216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8117..8136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8370..8387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8504..8525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8540..8545,8546..8563,8564..8570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8578..8594
                     /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     8583..8605
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8602..8618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8602..8618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8639..8657
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaaccaaaat caaatcacgg tatggcattt ctttacatat
       61 tctcaatacc tgactgacat agtatgcgct acatagtttt gtcctatttt ttttacaccg
      121 cgatttgaag tctaaagcca gagaatgagg aaaaacgaaa caaactgaga tcatttgtat
      181 aaggggagtc actgtacagc tgccaatacc aattttcctt ctgtcaaatc aaaaaagtat
      241 ccgcatacac atacacccct ccctctacaa tcttaattat ttactgattc ctttcgtatt
      301 tgtccagtat gagcaatact aagcatacca cttctcatca catggaactc aaaaggatca
      361 ttatccttac acttttattc atactcataa tgttgatatt tcgaaactca gtgtccttca
      421 aaatgacatt tcaagaacta ctgccacgat tttacaaaaa gaattcaaac tcagttagta
      481 ataataacag gccctcatct attttctcgg aaaacttggt ggattttgat gatgttaaca
      541 tggtcgataa gaccagactg tttatttttt tatttttcag tttcatcatt actataccgt
      601 ttatggtata aattagaaaa gttaaagcag catgttttca tttgaaacaa atactaatgc
      661 aataataggt acaccatcta ctacataatt aattgatagt ttccttttcc gttaagtgca
      721 tgcataggac gcccataatt ttttaagtta aaaaagcatg tatttaccta ttcgggaaat
      781 ttacatgaca tggatgccat aaggaacgaa aatgaaacat gcatgtcaga ggaaaaagaa
      841 caaagcgaga ggaaaaagct gttgcattac cacgtaattt tgtatataaa tatctgataa
      901 ataaccattt tatttccatc cacttcttct gtcgttcatc cgttcattga catcactaga
      961 gacaccagcg agctatcatc tgccagatct gtttagcttg ccttgtcccc gccgggtcac
     1021 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
     1081 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
     1141 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
     1201 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
     1261 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
     1321 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta
     1381 aggaaaagac tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg
     1441 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
     1501 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1561 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1621 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa
     1681 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1741 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     1801 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     1861 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
     1921 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     1981 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     2041 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     2101 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2161 tgatgctcga tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa
     2221 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     2281 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     2341 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     2401 ctaacgccgc catccagtgt cgaaaacggt accgggcccc cacacaccat agcttcaaaa
     2461 tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac
     2521 ttcaaaacac ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt
     2581 aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg
     2641 tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattt ttttttttga
     2701 tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc
     2761 tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag
     2821 aaagaaagca tagcaatcta atctaagttt taattacaac tcgagatggt tagcaaaggc
     2881 gaggaagata acatggctat aatcaaagag tttatgagat tcaaagtaca catggagggt
     2941 tcagtgaatg gtcatgaatt tgaaattgaa ggcgaaggcg agggcagacc ttacgaagga
     3001 actcaaacag caaaacttaa ggtaacaaaa ggtggtcctc tgccattcgc ctgggacatt
     3061 ctcagtccac aattcatgta cggttctaaa gcgtacgtca aacatccagc agacattcca
     3121 gattacttga aattgtcttt tccagaaggc tttaagtggg aaagagttat gaacttcgag
     3181 gatggagggg ttgtgaccgt tacgcaagat tcctctttac aagatggtga gtttatctac
     3241 aaggtcaaat taagggggac taattttcct tcagacgggc cagtcatgca gaaaaagact
     3301 atgggatggg aagcctcttc agagagaatg tatcctgaag atggcgctct aaaaggagaa
     3361 atcaagcaaa gattgaagtt aaaggacgga ggtcattatg atgcagaagt aaaaacaacc
     3421 tataaagcta aaaagccagt tcaacttcct ggtgcctaca atgttaacat caagctagac
     3481 attacatccc ataatgaaga ttacactata gtggaacagt atgaacgtgc tgaaggtaga
     3541 cacagtacag gtggtatgga tgaactgtac aagtaagcgg ccgccaccgc ggtggagctc
     3601 taagcaaata gctaaattat atacgaatta atattatgat taagtgttta cgtgagtgcg
     3661 atatttttat tactatctta tacagttgta tatactctat aaaatgagtt gtctattaat
     3721 taacgcgatg aatgctttct gggtttacct ctccaacaac tctagtttac ttctcaatac
     3781 attcaattgt atttgatttg tcaatacttc atcattaatc aattctatag ttttgttttt
     3841 ctcgtttatt tccaaattta atgcatcaat tttattattc aatttgtcgt tgattttggt
     3901 taatgatttt atggtttgat ctctggcatt gattgtttgt gttagttttt cattattgat
     3961 aattaaatta tttaagttag ttatcaactc ggtgttttca agtttcaagt tttcaatttc
     4021 tttagagttt attagatttg tcaaagtttc tgaattgctt gattggtcct gtagaagagt
     4081 atttgttgtt gtggataatt gattcaattt ttgagacaat tgctggaagg cgttgaaata
     4141 tctagcatca atctcatggt ttttttcccg agagtctcgt agattcaatt gttttaatat
     4201 atcttgggac cactcttgat ttgaactcat ggaaattaaa ctgggtgttg tgttgtggtg
     4261 taatgattgt accccctttg cttataattg tgtggccatt tcacgtgcat caagtccgta
     4321 ctatcttcac cactataaac tcttataggt ggattattcg tttcgaatgc tggtggcagt
     4381 gccggtttga aaataatctt acctacccat gggtggtctt ccgttaacgg tttgatgcat
     4441 cttaatgagt ctttgatata tcttttcctt gaaactagtt gtgtcgccgg tgcccacttc
     4501 gaatcgttac tggattgatt cttatttcca cattttagtg acaatttctt tggaaaatca
     4561 tgatttaaac aaccagtcac ggaaaagctt ttcgtactga atattgtacc gtactcaacg
     4621 tccgcattct catagcgaga ttctctttca tccaaatata ccaatctatt gatgtcatca
     4681 tcctcaacat atccatcttc atcatcctct tcgtaattag catcgtcttc gtcttcgtca
     4741 tcatcttcat ctccatttac atcttcattg ccacggttta acttttcttc acgattcgta
     4801 cgctcagcag aattcgaaat agttgacctt cctgttccct cgtataccgg ttcaaaatta
     4861 ttaattttca acactgaccg atcacagcgc aaaggatcta tagggttgga attcgatcta
     4921 cttctgaagt tttcatggat ggatgtttgt gccgttaaaa cagaatgtgc tgtgggagaa
     4981 aaagtggaag atagaacaga agtattggta ttagtagtac taccattgtc atcgatttct
     5041 tcttgaacac tattcagttc cttcgatctc attttttgtc caatgtccat attagagttt
     5101 tttctcgaac tggagtctgt tttaaagaaa tttcttagtt ttgatgtaaa taagccacaa
     5161 aaatcagttt gagaactctc ccctgccagc tagcgtttaa acggccacgc tggcccccta
     5221 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5281 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5341 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     5401 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5461 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5521 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5581 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5641 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5701 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5761 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5821 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     5881 atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat tgaagctcta
     5941 atttgtgagt ttagtattag gatctctacc cttggcgaaa agtcctctgc caacaatgat
     6001 gatatctgat ccaccactta caacttcgtc gacggttctg tactgctgac ccaatatgcg
     6061 tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
     6121 cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
     6181 attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6241 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6301 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6361 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     6421 gtcatgataa taatggtttc ttagacgtcg gcgcgccgag gacgtcaggt ggcacttttc
     6481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6541 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6601 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6661 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6721 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6781 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6841 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6901 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6961 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7021 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7081 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7141 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7201 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7261 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7321 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7381 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7441 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7501 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7561 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7621 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7681 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7741 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7801 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7861 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7921 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7981 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8041 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8101 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8161 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8221 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8281 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8341 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8401 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8461 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     8521 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     8581 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctcggaa
     8641 ttaaccctca ctaaagggaa caaaagctg
//
