LOCUS       sequence-238773-        6033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN HIS3 CNB1mutEF2.
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: pTJK504
FEATURES             Location/Qualifiers
     source          1..6033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(602..618)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(603..619)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(647..665)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(686..702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(686..702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(699..721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    710..726
                     /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(734..740,741..758,759..764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    779..800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(917..934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1088..1676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1168..1187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1847..2638,2639..2707))
                     /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     2470..2489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2708..2812)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2842..3345
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3393..3411
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3449..3471)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3571..3590
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3737..3924
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3925..4587
                     /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(4847..5302)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4984..5005)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5196..5215
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5428..5450
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5442..5459
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5443..5459
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5466..5485
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5466..5484
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5517..5533
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5517..5533
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 atgaaattag atagagatag ctcagggtct attgataaaa atgaatttat gagcattcct
       61 ggcgtttcgt caaaccctct tgctggacgt ataatggagg ttttcgctgc tgataatagt
      121 ggggacgtgg attttcaaga gttcatcaca ggattatcca ttttcagtgg gcgtgggtcc
      181 aaggacgaaa agttaagatt cgccttcaaa atctacgaca ttgacaagga cggtttcata
      241 tccaatggtg agttgttcat cgtgttgaag attatggtag gttctaatct ggacgatgaa
      301 cagctgcaac agatagtaga taggacgata gtggaaaacg atagcgacgg cgacggacgt
      361 ttaagtttcg aggagtttaa gaatgctatc gaaaccacag aagtggccaa gagtctgaca
      421 ttgcaatacg atgtgtaaga ctaggggaca cttcattcat ttatggtatg ccaatatttt
      481 taagaaaaga agaataatac gcgatattgt tttttaagga aggaacgcac actcgcccag
      541 ttagagtgct gatgatatat acatatatat atgtatatgt aacaaacaat aagaagctta
      601 tcgataccgt cgacctcgag ggggggcccg gtaccagctt ttgttccctt tagtgagggt
      661 taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      721 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      781 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      841 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      901 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      961 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1021 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1081 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1141 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1201 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1261 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1321 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1381 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1441 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1501 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1561 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1621 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1681 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1741 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1801 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1861 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1921 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1981 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2041 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2101 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2161 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2221 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2281 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2341 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2401 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2461 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2521 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2581 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2641 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2701 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2761 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2821 cccgaaaagt gccacctgac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt
     2881 aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg
     2941 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca
     3001 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa
     3061 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt
     3121 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg
     3181 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg
     3241 aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta atttatatat
     3301 ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa ggaccctaag
     3361 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3421 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3481 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3541 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3601 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
     3661 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
     3721 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
     3781 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat
     3841 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag
     3901 caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa gcgtattaca
     3961 aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc gatagagcac
     4021 tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca atcgcaagtg
     4081 attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct ggccaagcat
     4141 tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga ccatcacacc
     4201 actgaagact gcgggattgc tctcggtcaa gcttttaaag aggccctagg ggccgtgcgt
     4261 ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg
     4321 gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta
     4381 ggagatctct cttgcgagat gatcccgcat tttcttgaaa gctttgcaga ggctagcaga
     4441 attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg
     4501 ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac caacgatgtt
     4561 ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagctgc agcatacgat
     4621 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta
     4681 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg
     4741 ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg
     4801 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     4861 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4921 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4981 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5041 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5101 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5161 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5221 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     5281 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     5341 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5401 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     5461 aattgtaata cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta
     5521 gaactagtgg atccacgcta tgaaaaagga aaacaaatga ctcttataat ttgtttccct
     5581 cgacttctct atttaaatcc aagattttaa ctaataaact atttaataat aaaacaatta
     5641 actctttata tggaagcctt ggaaattagc cgccaaaatg ggatatacat tccgtgcgaa
     5701 gtgaccgcgt ggaaggccgg gtatcatctt aaaaatcact agtttctttt ttagcggaat
     5761 gcaataaagg tgctttgtgc tggtgtggtt tacacggaac atctagtagc taaaacttgg
     5821 taactcaatg gtgatcagaa tccatagaag catttttatt tcttaaaatg ggtgctgctc
     5881 cttccaaaat tgtggatggt cttttagaag atacaaattg tatgtacact tcggagtgag
     5941 gaaaagaaag aaaggggaaa ttaaccgatt ttactaacac tgacactttg aacagttgat
     6001 agagatgaaa ttgaaaggtt aaggaagaga ttc
//
