LOCUS       sequence-306292-        7487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrating plasmid for pTETO1-Venus reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  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 7487)
  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: pJZC530
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(129..134,135..152,153..159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..183
                     /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     172..194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     191..207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        228..246
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2213..2231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2426..3139
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEGGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3141..3157)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3141..3157)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4292..4311)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4293..4311)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4318..4335)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4318..4334)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4327..4349)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4475..4930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4562..4581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4772..4793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5286..5305)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5405..5427
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5465..5483)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5568..5672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5673..5741,5742..6533)
                     /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(5891..5910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6704..7292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7193..7212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7446..7463
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag ctatgaccat gattacgcca agctcggaat taaccctcac
      241 taaagggaac aaaagctggg ccggccattt aaatgtttaa acggccacgc tggccatatt
      301 tctaccaatc tctcaacact gagtaatggt agttataaga aagagaccga gttagggaca
      361 gttagaggcg gtggagatat tccttatggc atgtctggcg atgataaaac ttttcaaacg
      421 gcagccccga tctaaaagag ctgacaggga aatggtcaga aaaagaaacg tgcacccgcc
      481 cgtctggacg cgccgctcac ccgcacggca gagaccaatc agtaaaaatc aacggttaac
      541 gacattacta tatatataat ataggaagca tttaatagaa cagcatcgta atatatgtgt
      601 actttgcagt tatgacgcca gatggcagta gtggaagata ttctttattg aaaaatagct
      661 tgtcacctta cgtacaatct tgatccggag cccgtcgctt tgagagaaat cagaagattc
      721 caaaagtcta ctgaactttt gatcagaaag ttgcctttcc aaagactagt cagagaaatc
      781 gcccaagatt tcaagaccga tctaagattc caatcctctg ccatcggtgc tctacaggaa
      841 tccgttgaag cttacttggt ctctttgttc gaagacacca acttggccgc tatccacgct
      901 aagcgtgtta ccatccaaaa gaaggatatc aagttggcta gaagattgag aggtgaaaga
      961 tcctaagcac acagggattg ttgtacaatt actttttatg attttagata cacacaaatc
     1021 ttttctacga ttacctttca tttattttcg tttcatgttt tattaatcat tctttttgta
     1081 tatttcacat cgggtggggt ttttatatta aattttaata acataatctt ttaaagtttc
     1141 tactttctat cctgtatatt atttccatat ctatatctat attttcaagc ttgttcacat
     1201 ccttatcgga attgttccag cgtttttgta cagaaaatga tgttatggta acgagctata
     1261 tcagacctac tagtcttata gagctgtaca gagtcttgga aacataacca acacaccaac
     1321 aagatgtcat ttgattcgtt actcgacaag aatgataagc tggtaaaagt ttgcgggatt
     1381 caaaccgtcg aggctgccga aactgcgctt caagcaggcg ctgatttgat agggatcata
     1441 tgtgtcccca acaggaagcg gactatcgag agcgctgtgg ctcgtgaaat atccaaattg
     1501 attcacaaat caggtactac aaagctggtg ggggtgttca ggaatcaatc tgttgaggac
     1561 gtacatcggc tttctgagga atatgacctt gacataatcc aattacatgg tgatgaatca
     1621 tggccagagt actataacgt cattaagaaa ccaataatca aaagagtcat attccctaga
     1681 gatgtcgatg ttgtaacaca agtgtgtcaa agaaaaacct tggtatgtct accattgttc
     1741 gactccgagg caggcggtac aggtgaaaag ctggactggt ccagtatttc ctcctgggca
     1801 agcgaacaga ataacgtatt ttatatacta gcgggtggac taactgcaga aaacgtcatg
     1861 gaggcggtgg ggttgcctgg tgtaattggt gttgacgtct caggtggtgt agagactgac
     1921 ggcgtcaaag acaatgacaa gataattaaa tatgtacaaa atgcaaagaa acaatgaatt
     1981 acgaagagaa gtctcaatga ggttttacaa acaatcctat aatacatata tgccacttag
     2041 atatactttg aatactgaat atactgactc attatattct ttaattgtag tttctcaaaa
     2101 atattcatac aggttagcta tgaattcatg ctataatttc cattttgttc atgtaagata
     2161 cacataggga taaaatgtga taactaatca gcggtaccgg gccctttacc actccctatc
     2221 agtgatagag aaaagtgaaa gtcgagctcg gtaccctatg gcatgcatgt gctctgtatg
     2281 tatataaaac tcttgttttc ttcttttctc taaatattct ttccttatac attaggtcct
     2341 ttgtagcata aattactata cttctataga cacgcaaaca caaatacaca cactaaatta
     2401 cccggatcaa ttcgggatgc tcgagtctaa aggtgaagaa ttattcactg gtgttgtccc
     2461 aattttggtt gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg
     2521 tgaaggtgat gctacttacg gtaaattgac cttaaaattg atttgtacta ctggtaaatt
     2581 gccagttcca tggccaacct tagtcactac tttaggttat ggtttgcaat gttttgctag
     2641 atacccagat catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt
     2701 tcaagaaaga actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa
     2761 gtttgaaggt gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaagg
     2821 tggtaacatt ttaggtcaca aattggaata caactataac tctcacaatg tttacatcac
     2881 tgctgacaaa caaaagaatg gtatcaaagc taacttcaaa attagacaca acattgaaga
     2941 tggtggtgtt caattagctg accattatca acaaaatact ccaattggtg atggtccagt
     3001 cttgttacca gacaaccatt acttatccta tcaatctgcc ttatccaaag atccaaacga
     3061 aaagagagac cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat
     3121 ggatgaattg tacaaataag gatccactag ttctagacac ctgcaacatg cgtgagtaaa
     3181 taagcggccg ccaccgcggt ggagctctaa gcaaatagct aaattatata cgaattaata
     3241 ttatgattaa gtgtttacgt gagtgcgata tttttattac tatcttatac agttgtatat
     3301 actctataaa atgagttgtc tattaattaa cgcgatgaat gctttctggg tttacctctc
     3361 caacaactct agtttacttc tcaatacatt caattgtatt tgatttgtca atacttcatc
     3421 attaatcaat tctatagttt tgtttttctc gtttatttcc aaatttaatg catcaatttt
     3481 attattcaat ttgtcgttga ttttggttaa tgattttatg gtttgatctc tggcattgat
     3541 tgtttgtgtt agtttttcat tattgataat taaattattt aagttagtta tcaactcggt
     3601 gttttcaagt ttcaagtttt caatttcttt agagtttatt agatttgtca aagtttctga
     3661 attgcttgat tggtcctgta gaagagtatt tgttgttgtg gataattgat tcaatttttg
     3721 agacaattgc tggaaggcgt tgaaatatct agcatcaatc tcatggtttt tttcccgaga
     3781 gtctcgtaga ttcaattgtt ttaatatatc ttgggaccac tcttgatttg aactcatgga
     3841 aattaaactg ggtgttgtgt tgtggtgtaa tgattgtacc ccctttgctt ataattgtgt
     3901 ggttccgatg ctgacttact gggtattata tgtgtgccca atagaaagag aacaattgac
     3961 ccggttattg caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact
     4021 ccgaaatact tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc
     4081 aatgattacg gcattgatat cgtccaactg catggagatg agtcgtggcg agaataccaa
     4141 gagttcctcg gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta
     4201 ctcagtgcag cttcacagaa acctcattcg tttattccct tgtttgattc agaaggccgg
     4261 ccatttaaat gtttaaacgg ccacgctggc cccctatagt gagtcgtatt acaattcact
     4321 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4381 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4441 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     4501 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4561 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4621 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4681 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4741 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4801 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4861 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4921 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4981 tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tattaggatc
     5041 tctacccttg gcgaaaagtc ctctgccaac aatgatgata tctgatccac cacttacaac
     5101 ttcgtcgacg gttctgtact gctgacccaa tatgcgtata tataccaatc taagtctgtg
     5161 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa
     5221 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt atggtgcact
     5281 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5341 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5401 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5461 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5521 acgtcggcgc gccgaggacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5581 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5641 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5701 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5761 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5821 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5881 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5941 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6001 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6061 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6121 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6181 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6241 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6301 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6361 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6421 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6481 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6541 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6601 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6661 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6721 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6781 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6841 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6901 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6961 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7021 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7081 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7141 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7201 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7261 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7321 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7381 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7441 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgc
//
