LOCUS       sequence-238889-        7710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN TRP1 CNA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7710)
  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 7710)
  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: pTJK517
FEATURES             Location/Qualifiers
     source          1..7710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2052..2070)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2091..2107)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2091..2107)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2104..2126)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2115..2131
                     /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(2139..2145,2146..2163,2164..2169))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2184..2205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2322..2339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2493..3081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2573..2592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3252..4043,4044..4112))
                     /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     3875..3894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4113..4217)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4254..4757
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4798..4816
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4854..4876)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4976..4995
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5016..5296
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5297..5971
                     /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(6074..6529)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6211..6232)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6423..6442
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6655..6677
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6669..6686
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6670..6686
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6693..6712
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6693..6711
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     6744..6760
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6744..6760
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 atgtcgaaag acttgaattc ttcacgcctc aaaatcatta aacctaatga ctcttacata
       61 aaggttgacc ggaaaaaaga tttgacaaaa tacgaattag aaaacggtaa agtaatttct
      121 actaaggacc gacctatcgc ttctgtacct gccataacag gaaagatacc aagtgatgag
      181 gaagtattcg actccaagac gggattacct aatcattcct ttttaagaga gcatttcttt
      241 catgagggtc gactttctaa ggaacaggcc ataaaaatct taaatatgtc aactgtagca
      301 ttgagtaaag aacccaatct actaaaactc aaagcgccaa ttactatatg tggtgatatt
      361 cacgggcagt attatgattt attgaaactg tttgaagttg gcggtgaccc cgccgaaatc
      421 gactatttat tcttggggga ttatgttgat agaggtgcat tctcttttga gtgtctgatt
      481 tatttgtact ccttgaagtt gaataattta gggagatttt ggatgctaag aggtaaccat
      541 gagtgtaagc acttgacctc ttattttact tttaagaatg agatgttgca caaatacgat
      601 atggaagttt acgatgcttg ctgcagatca ttcaatgtct taccattagc agctttaatg
      661 aacggacaat atttttgtgt gcatggtggt atctctccag agttaaaatc agtagaggat
      721 gttaataaaa ttaatagatt tcgagaaatc ccatctcgtg gtctcatgtg tgacctacta
      781 tgggccgatc ctgtcgaaaa ttatgatgat gcaagagatg gtagcgaatt tgatcagagc
      841 gaggatgaat tcgtacctaa cagtttgagg ggttgctctt tcgccttcac ttttaaagca
      901 tcatgcaagt ttttgaaggc aaatggtttg ttatctatta ttagagcaca cgaagcacag
      961 gatgctgggt acagaatgta taaaaacaat aaagtaacag gcttcccgag cttaataacc
     1021 atgttcagtg cgccaaacta cctggacaca tatcataata aagctgctgt gttaaaatat
     1081 gaagaaaacg tcatgaacat caggcagttt cacatgtctc cgcaccctta ctggttgcct
     1141 gattttatgg atgttttcac ctggtcacta ccttttgttg gcgaaaaagt tactagcatg
     1201 ttagtgtcta tattaaacat atgtagtgag caggaacttg acccagaatc ggaacccaaa
     1261 gctgcggagg agactgtaaa ggcaagagca aacgcaacta aggagaccgg caccccatct
     1321 gatgaaaagg cgtcatcagc gatattagaa gatgaaaccc gaagaaaggc tttgagaaat
     1381 aagatattag ctattgctaa agtttcaaga atgttttcgg tgctaagaga agagagcgaa
     1441 aaagtggaat atttgaaaac tatgaatgcc ggtgtcttac ctcgtggtgc tctagctcgt
     1501 gggactgaag gtttgaatga aacgctaagc acttttgaaa aggctagaaa ggaagacctt
     1561 attaatgaaa aattaccacc atctttatcg gaggttgaac aagagaagat taaatactac
     1621 gaaaaaatat taaagggagc ggagaaaaag ccacaactgt gataaatctc attttattct
     1681 tttttattcc tataagatgc aacatcgaat tcttgtgcta ctgtcacgat ccgcaagaga
     1741 taatatgaac taacaaaata cgaagaaaat atagatcatt tccagtctat ttcttgattt
     1801 ccatacttaa ctgcgagcag aataaatata aacacatatt ttttagtttt attttattat
     1861 taatgaaccc tagttaccaa ttagttatgt aaacgctttt cgtaagaatg taagttatga
     1921 acaccacctt ctagttgagc agaagcggtt ctaaattcaa atctaacttt accgctttgg
     1981 acattttcct ttaataaagt tagggacttg taaccaatgc tcgagggggg gcccggtacc
     2041 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
     2101 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2161 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2221 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2281 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3241 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3301 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3361 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3421 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3481 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3541 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3601 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3661 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3721 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3781 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3841 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3901 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3961 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4021 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4081 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4141 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4201 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt
     4261 gctataaaaa taattataat ttaaattttt taatataaat atataaatta aaaatagaaa
     4321 gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag
     4381 ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc
     4441 gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt
     4501 tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat
     4561 atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt
     4621 ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata
     4681 aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa
     4741 gttacaggca agcgatccgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4801 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4861 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4921 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     4981 tcagagcaga ttgtactgag agtgcaccat agatcacgac attactatat atataatata
     5041 ggaagcattt aatagaacag catcgtaata tatgtgtact ttgcagttat gacgccagat
     5101 ggcagtagtg gaagatattc tttattgaaa aatagcttgt caccttacgt acaatcttga
     5161 tccggagctt ttcttttttt gccgattaag aattaattcg gtcgaaaaaa gaaaaggaga
     5221 gggccaagag ggagggcatt ggtgactatt gagcacgtga gtatacgtga ttaagcacac
     5281 aaaggcagct tggagtatgt ctgttattaa tttcacaggt agttctggtc cattggtgaa
     5341 agtttgcggc ttgcagagca cagaggccgc agaatgtgct ctagattccg atgctgactt
     5401 gctgggtatt atatgtgtgc ccaatagaaa gagaacaatt gacccggtta ttgcaaggaa
     5461 aatttcaagt cttgtaaaag catataaaaa tagttcaggc actccgaaat acttggttgg
     5521 cgtgtttcgt aatcaaccta aggaggatgt tttggctctg gtcaatgatt acggcattga
     5581 tatcgtccaa ctgcatggag atgagtcgtg gcaagaatac caagagttcc tcggtttgcc
     5641 agttattaaa agactcgtat ttccaaaaga ctgcaacata ctactcagtg cagcttcaca
     5701 gaaacctcat tcgtttattc ccttgtttga ttcagaagca ggtgggacag gtgaactttt
     5761 ggattggaac tcgatttctg actgggttgg aaggcaagag agccccgaaa gcttacattt
     5821 tatgttagct ggtggactga cgccagaaaa tgttggtgat gcgcttagat taaatggcgt
     5881 tattggtgtt gatgtaagcg gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat
     5941 agcaaatttc gtcaaaaatg ctaagaaata ggttattact gagtagtatt tatttaagta
     6001 ttgtttgtgc acttgccgat ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6061 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     6121 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6181 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6241 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6301 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6361 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6421 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6481 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     6541 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6601 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     6661 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
     6721 agctccaccg cggtggcggc cgctctagaa ctagtggatc cgtcttcgtc aacaattggt
     6781 tcaggtttag acgatactaa gtcaggtaca acctcgtcca tctcttcatc ttcatcttcg
     6841 tcatcatagt cctcttcatc ttcatcgtct tcatcatcat caccctcaat atgcacaact
     6901 ctttttgatt ttttagttct ctgcttttct tgccattcca aatacaattt ttctacctta
     6961 ttataattta aattgaaaca gtcgttatat atgttgatga cctctacggc aatcggtaaa
     7021 gccgagtcgt cttcaacatt agtttcaaat tcatctatca ttgcataaag taacgtttct
     7081 tcgatcaaag cggcgtcgac aactttttca tttttgaaaa ggtctactac aatccctgta
     7141 atccagtctc tcttctcagc tgagtctgga ccaccccaac tgttttctac ggcaacatcc
     7201 aacgcatccc acttataaat aaccatggaa acaccgagct caaaacgtgc ctgttgcttt
     7261 tcgtcagaga acattagatt ggttttgcct tgctcagcct gaacaaatgc tgtctcatca
     7321 atatattgtg tgctcattgt tccgtaagga ttatcactat aaaaattttc cgcagatttc
     7381 ttttggactc cacctttctt cctgttgtaa taaatacaaa ctatttgaat caatttgata
     7441 ttcatctata tctatttcaa aatttttcat gtcatcgcct cttgaaacgt gaattttcca
     7501 attctgaaaa agaacgtact actgggaaac aaaagggaaa aatgtataaa tcctttaatg
     7561 tttttaaatc aagaggcatt attataaaag aacgaagcaa agcctttaat atttgcttta
     7621 ttaaaggtac tattcaaaga aaagtttttt tagattcttt tttttttttt cacgtattag
     7681 ctcagctgcc ataaaacact ctcaacgcca
//
