LOCUS       sequence-238925-        7435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN TRP1 CNA1-Delta-CBD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7435)
  AUTHORS   Connolly S, Quasi-Woode D, Waldron L, Eberly C, Waters K, Muller EM,
            Kingsbury TJ
  TITLE     Calcineurin Regulatory Subunit Calcium-Binding Domains 
            Differentially Contribute to Calcineurin Signaling in Saccharomyces 
            cerevisiae.
  JOURNAL   Genetics. 2018 Jul;209(3):801-813. doi: 10.1534/genetics.118.300911.
            Epub 2018 May 7.
  PUBMED    29735720
REFERENCE   2  (bases 1 to 7435)
  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: 
            "10.1534/genetics.118.300911"; journalName: "Genetics"; date: 
            "2018-07"; volume: "209"; issue: "3"; pages: "801-813"
COMMENT     Sequence Label: pTJK519
FEATURES             Location/Qualifiers
     source          1..7435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1624..1642)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1663..1679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1663..1679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1676..1698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1687..1703
                     /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)."
     promoter        complement(join(1711..1717,1718..1735,1736..1741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1756..1777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1894..1911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2065..2653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2145..2164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2824..3615,3616..3684))
                     /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     3447..3466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3685..3789)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3826..4329
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4370..4388
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4426..4448)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4548..4567
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4588..4868
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             4869..5543
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(5646..6101)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5783..5804)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5995..6014
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6227..6249
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6241..6258
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6242..6258
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6265..6284
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6265..6283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     6316..6332
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6316..6332
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 ataacaggaa agataccaag tgatgaggaa gtattcgact ccaagacggg attacctaat
       61 cattcctttt taagagagca tttctttcat gagggtcgac tttctaagga acaggccata
      121 aaaatcttaa atatgtcaac tgtagcattg agtaaagaac ccaatctact aaaactcaaa
      181 gcgccaatta ctatatgtgg tgatattcac gggcagtatt atgatttatt gaaactgttt
      241 gaagttggcg gtgaccccgc cgaaatcgac tatttattct tgggggatta tgttgataga
      301 ggtgcattct cttttgagtg tctgatttat ttgtactcct tgaagttgaa taatttaggg
      361 agattttgga tgctaagagg taaccatgag tgtaagcact tgacctctta ttttactttt
      421 aagaatgaga tgttgcacaa atacgatatg gaagtttacg atgcttgctg cagatcattc
      481 aatgtcttac cattagcagc tttaatgaac ggacaatatt tttgtgtgca tggtggtatc
      541 tctccagagt taaaatcagt agaggatgtt aataaaatta atagatttcg agaaatccca
      601 tctcgtggtc tcatgtgtga cctactatgg gccgatcctg tcgaaaatta tgatgatgca
      661 agagatggta gcgaatttga tcagagcgag gatgaattcg tacctaacag tttgaggggt
      721 tgctctttcg ccttcacttt taaagcatca tgcaagtttt tgaaggcaaa tggtttgtta
      781 tctattatta gagcacacga agcacaggat gctgggtaca gaatgtataa aaacaataaa
      841 gtaacaggct tcccgagctt aataaccatg ttcagtgcgc caaactacct ggacacatat
      901 cataataaag ctgctgtgtt aaaatatgaa gaaaacgtca tgaacatcag gcagtttcac
      961 atgtctccgc acccttactg gttgcctgat tttatggatg ttttcacctg gtcactacct
     1021 tttgttggcg aaaaagttac tagcatgtta gtgtctatat taaacatatg tagtgagcag
     1081 gaacttgacc cagaatcgga acccaaagct gcggaggaga ctgtaaaggc aagagcaaac
     1141 gcaactaagg agaccggcac cccatctgat gaaaaggcgt catcagcgat attagaagat
     1201 gaaacccaac tgtgataaat ctcattttat tcttttttat tcctataaga tgcaacatcg
     1261 aattcttgtg ctactgtcac gatccgcaag agataatatg aactaacaaa atacgaagaa
     1321 aatatagatc atttccagtc tatttcttga tttccatact taactgcgag cagaataaat
     1381 ataaacacat attttttagt tttattttat tattaatgaa ccctagttac caattagtta
     1441 tgtaaacgct tttcgtaaga atgtaagtta tgaacaccac cttctagttg agcagaagcg
     1501 gttctaaatt caaatctaac tttaccgctt tggacatttt cctttaataa agttagggac
     1561 ttgtaaccaa tgctcgaatc ttgctgaaaa actcgagggg gggcccggta ccagcttttg
     1621 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     1681 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1741 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1801 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1861 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1921 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1981 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2041 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2101 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2161 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2221 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2281 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2341 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2401 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2461 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2521 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2581 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2641 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2701 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2761 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2821 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2881 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2941 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3001 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3061 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3121 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3181 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3241 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3301 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3361 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3421 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3481 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3541 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3601 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3661 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3721 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3781 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa
     3841 aataattata atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa
     3901 agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg
     3961 ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt
     4021 tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc
     4081 gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag
     4141 tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact
     4201 cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca
     4261 cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg
     4321 caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4381 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4441 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4501 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4561 gattgtactg agagtgcacc atagatcacg acattactat atatataata taggaagcat
     4621 ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag
     4681 tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc
     4741 ttttcttttt ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag
     4801 agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag
     4861 cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg
     4921 gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta
     4981 ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa
     5041 gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc
     5101 gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc
     5161 aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta
     5221 aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc
     5281 attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga
     5341 actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag
     5401 ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg
     5461 ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt
     5521 tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt
     5581 gcacttgccg atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     5641 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5701 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5761 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5821 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5881 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5941 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     6001 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     6061 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     6121 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6181 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6241 tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt ggagctccac
     6301 cgcggtggcg gccgctctag aactagtgga tccgtcttcg tcaacaattg gttcaggttt
     6361 agacgatact aagtcaggta caacctcgtc catctcttca tcttcatctt cgtcatcata
     6421 gtcctcttca tcttcatcgt cttcatcatc atcaccctca atatgcacaa ctctttttga
     6481 ttttttagtt ctctgctttt cttgccattc caaatacaat ttttctacct tattataatt
     6541 taaattgaaa cagtcgttat atatgttgat gacctctacg gcaatcggta aagccgagtc
     6601 gtcttcaaca ttagtttcaa attcatctat cattgcataa agtaacgttt cttcgatcaa
     6661 agcggcgtcg acaacttttt catttttgaa aaggtctact acaatccctg taatccagtc
     6721 tctcttctca gctgagtctg gaccacccca actgttttct acggcaacat ccaacgcatc
     6781 ccacttataa ataaccatgg aaacaccgag ctcaaaacgt gcctgttgct tttcgtcaga
     6841 gaacattaga ttggttttgc cttgctcagc ctgaacaaat gctgtctcat caatatattg
     6901 tgtgctcatt gttccgtaag gattatcact ataaaaattt tccgcagatt tcttttggac
     6961 tccacctttc ttcctgttgt aataaataca aactatttga atcaatttga tattcatcta
     7021 tatctatttc aaaatttttc atgtcatcgc ctcttgaaac gtgaattttc caattctgaa
     7081 aaagaacgta ctactgggaa acaaaaggga aaaatgtata aatcctttaa tgtttttaaa
     7141 tcaagaggca ttattataaa agaacgaagc aaagccttta atatttgctt tattaaaggt
     7201 actattcaaa gaaaagtttt tttagattct tttttttttt ttcacgtatt agctcagctg
     7261 ccataaaaca ctctcaacgc caatgtcgaa agacttgaat tcttcacgcc tcaaaatcat
     7321 taaacctaat gactcttaca taaaggttga ccggaaaaaa gatttgacaa aatacgaatt
     7381 agaaaacggt aaagtaattt ctactaagga ccgacctatc gcttctgtac ctgcc
//
