LOCUS       sequence-238896-        6033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN HIS3 CNB1mutEF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6033)
  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 6033)
  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: pTJK506
FEATURES             Location/Qualifiers
     source          1..6033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(164..180)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(165..181)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(209..227)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(248..264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(248..264)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(261..283)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    272..288
                     /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(296..302,303..320,321..326))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    341..362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(479..496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(650..1238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(730..749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1409..2200,2201..2269))
                     /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     2032..2051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2270..2374)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2404..2907
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2955..2973
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3011..3033)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3133..3152
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3299..3486
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3487..4149
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(4409..4864)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4546..4567)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4758..4777
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4990..5012
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5004..5021
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5005..5021
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5028..5047
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5028..5046
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5079..5095
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5079..5095
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gactagggga cacttcattc atttatggta tgccaatatt tttaagaaaa gaagaataat
       61 acgcgatatt gttttttaag gaaggaacgc acactcgccc agttagagtg ctgatgatat
      121 atacatatat atatgtatat gtaacaaaca ataagaagct tatcgatacc gtcgacctcg
      181 agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
      241 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      301 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      361 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      421 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      481 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1381 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1441 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1501 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1561 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1621 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1681 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1741 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1801 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1861 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1921 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1981 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2041 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2101 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2161 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2221 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2281 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2341 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2401 acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg
     2461 gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa
     2521 agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca
     2581 acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta aatagatata
     2641 cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca
     2701 cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt
     2761 agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt
     2821 tctttaattt ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa
     2881 attataatta tttttatagc acgtgatgaa aaggacccta agaaaccatt attatcatga
     2941 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     3001 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     3061 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     3121 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa ttcccgtttt
     3181 aagagcttgg tgagcgctag gagtcactgc caggtatcgt ttgaacacgg cattagtcag
     3241 ggaagtcata acacagtcct ttcccgcaat tttctttttc tattactctt ggcctcctct
     3301 agtacactct atattttttt atgcctcggt aatgattttc attttttttt ttccacctag
     3361 cggatgactc tttttttttc ttagcgattg gcattatcac ataatgaatt atacattata
     3421 taaagtaatg tgatttcttc gaagaatata ctaaaaaatg agcaggcaag ataaacgaag
     3481 gcaaagatga cagagcagaa agccctagta aagcgtatta caaatgaaac caagattcag
     3541 attgcgatct ctttaaaggg tggtccccta gcgatagagc actcgatctt cccagaaaaa
     3601 gaggcagaag cagtagcaga acaggccaca caatcgcaag tgattaacgt ccacacaggt
     3661 atagggtttc tggaccatat gatacatgct ctggccaagc attccggctg gtcgctaatc
     3721 gttgagtgca ttggtgactt acacatagac gaccatcaca ccactgaaga ctgcgggatt
     3781 gctctcggtc aagcttttaa agaggcccta ggggccgtgc gtggagtaaa aaggtttgga
     3841 tcaggatttg cgcctttgga tgaggcactt tccagagcgg tggtagatct ttcgaacagg
     3901 ccgtacgcag ttgtcgaact tggtttgcaa agggagaaag taggagatct ctcttgcgag
     3961 atgatcccgc attttcttga aagctttgca gaggctagca gaattaccct ccacgttgat
     4021 tgtctgcgag gcaagaatga tcatcaccgt agtgagagtg cgttcaaggc tcttgcggtt
     4081 gccataagag aagccacctc gcccaatggt accaacgatg ttccctccac caaaggtgtt
     4141 cttatgtagt gacaccgatt atttaaagct gcagcatacg atatatatac atgtgtatat
     4201 atgtatacct atgaatgtca gtaagtatgt atacgaacag tatgatactg aagatgacaa
     4261 ggtaatgcat cattctatac gtgtcattct gaacgaggcg cgctttcctt ttttcttttt
     4321 gctttttctt tttttttctc ttgaactcga cggatctatg cggtgtgaaa taccgcacag
     4381 atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc
     4441 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4501 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4561 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4621 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4681 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4741 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4801 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4861 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4921 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     4981 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
     5041 tatagggcga attggagctc caccgcggtg gcggccgctc tagaactagt ggatccacgc
     5101 tatgaaaaag gaaaacaaat gactcttata atttgtttcc ctcgacttct ctatttaaat
     5161 ccaagatttt aactaataaa ctatttaata ataaaacaat taactcttta tatggaagcc
     5221 ttggaaatta gccgccaaaa tgggatatac attccgtgcg aagtgaccgc gtggaaggcc
     5281 gggtatcatc ttaaaaatca ctagtttctt ttttagcgga atgcaataaa ggtgctttgt
     5341 gctggtgtgg tttacacgga acatctagta gctaaaactt ggtaactcaa tggtgatcag
     5401 aatccataga agcattttta tttcttaaaa tgggtgctgc tccttccaaa attgtggatg
     5461 gtcttttaga agatacaaat tgtatgtaca cttcggagtg aggaaaagaa agaaagggga
     5521 aattaaccga ttttactaac actgacactt tgaacagttg atagagatga aattgaaagg
     5581 ttaaggaaga gattcatgaa attagataga gatagctcag ggtctattga taaaaatgaa
     5641 tttatgagca ttcctggcgt ttcgtcaaac cctcttgctg gacgtataat ggaggttttc
     5701 gatgctgata atagtgggga cgtggatttt caagagttca tcacaggatt atccattttc
     5761 agtgggcgtg ggtccaagga cgaaaagtta agattcgcct tcaaaatcta cgacattgac
     5821 aaggacggtt tcatatccaa tggtgagttg ttcatcgtgt tgaagattat ggtaggttct
     5881 aatctggacg atgaacagct gcaacagata gtagatagga cgatagtgga aaacgctagc
     5941 gacggcgacg gacgtttaag tttcgaggag tttaagaatg ctatcgaaac cacagaagtg
     6001 gccaagagtc tgacattgca atacgatgtg taa
//
