LOCUS       sequence-238920-        6033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN HIS3 CNB1mutEF1.
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: pTJK503
FEATURES             Location/Qualifiers
     source          1..6033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     75..91
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     75..91
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1193..1209)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1194..1210)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1238..1256)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1277..1293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1277..1293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1290..1312)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1301..1317
                     /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(1325..1331,1332..1349,1350..1355))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1370..1391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1508..1525)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1679..2267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1759..1778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2438..3229,3230..3298))
                     /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     3061..3080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3299..3403)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3433..3936
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3984..4002
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4040..4062)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4162..4181
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4328..4515
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             4516..5178
                     /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(5438..5893)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5575..5596)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5787..5806
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6019..6033,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6033,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gagctccacc
       61 gcggtggcgg ccgctctaga actagtggat ccacgctatg aaaaaggaaa acaaatgact
      121 cttataattt gtttccctcg acttctctat ttaaatccaa gattttaact aataaactat
      181 ttaataataa aacaattaac tctttatatg gaagccttgg aaattagccg ccaaaatggg
      241 atatacattc cgtgcgaagt gaccgcgtgg aaggccgggt atcatcttaa aaatcactag
      301 tttctttttt agcggaatgc aataaaggtg ctttgtgctg gtgtggttta cacggaacat
      361 ctagtagcta aaacttggta actcaatggt gatcagaatc catagaagca tttttatttc
      421 ttaaaatggg tgctgctcct tccaaaattg tggatggtct tttagaagat acaaattgta
      481 tgtacacttc ggagtgagga aaagaaagaa aggggaaatt aaccgatttt actaacactg
      541 acactttgaa cagttgatag agatgaaatt gaaaggttaa ggaagagatt catgaaatta
      601 gctagagata gctcagggtc tattgataaa aatgaattta tgagcattcc tggcgtttcg
      661 tcaaaccctc ttgctggacg tataatggag gttttcgatg ctgataatag tggggacgtg
      721 gattttcaag agttcatcac aggattatcc attttcagtg ggcgtgggtc caaggacgaa
      781 aagttaagat tcgccttcaa aatctacgac attgacaagg acggtttcat atccaatggt
      841 gagttgttca tcgtgttgaa gattatggta ggttctaatc tggacgatga acagctgcaa
      901 cagatagtag ataggacgat agtggaaaac gatagcgacg gcgacggacg tttaagtttc
      961 gaggagttta agaatgctat cgaaaccaca gaagtggcca agagtctgac attgcaatac
     1021 gatgtgtaag actaggggac acttcattca tttatggtat gccaatattt ttaagaaaag
     1081 aagaataata cgcgatattg ttttttaagg aaggaacgca cactcgccca gttagagtgc
     1141 tgatgatata tacatatata tatgtatatg taacaaacaa taagaagctt atcgataccg
     1201 tcgacctcga gggggggccc ggtaccagct tttgttccct ttagtgaggg ttaatttcga
     1261 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1321 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1381 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1441 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1501 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1561 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1861 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2101 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2341 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2401 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2461 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2521 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2581 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2641 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2701 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2761 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2821 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2881 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2941 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3001 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3061 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3121 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3181 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3241 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3301 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3361 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3421 tgccacctga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt taatgatgga
     3481 ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg gattttagaa
     3541 agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac aaaggtttaa
     3601 aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa agcagattaa
     3661 atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg
     3721 tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag gaagagcaag
     3781 ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa
     3841 actatttttt ctttaatttc tttttttact ttctattttt aatttatata tttatattaa
     3901 aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggaccctaa gaaaccatta
     3961 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     4021 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4081 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4141 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataaat
     4201 tcccgtttta agagcttggt gagcgctagg agtcactgcc aggtatcgtt tgaacacggc
     4261 attagtcagg gaagtcataa cacagtcctt tcccgcaatt ttctttttct attactcttg
     4321 gcctcctcta gtacactcta tattttttta tgcctcggta atgattttca tttttttttt
     4381 tccacctagc ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta
     4441 tacattatat aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga
     4501 taaacgaagg caaagatgac agagcagaaa gccctagtaa agcgtattac aaatgaaacc
     4561 aagattcaga ttgcgatctc tttaaagggt ggtcccctag cgatagagca ctcgatcttc
     4621 ccagaaaaag aggcagaagc agtagcagaa caggccacac aatcgcaagt gattaacgtc
     4681 cacacaggta tagggtttct ggaccatatg atacatgctc tggccaagca ttccggctgg
     4741 tcgctaatcg ttgagtgcat tggtgactta cacatagacg accatcacac cactgaagac
     4801 tgcgggattg ctctcggtca agcttttaaa gaggccctag gggccgtgcg tggagtaaaa
     4861 aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt ggtagatctt
     4921 tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc
     4981 tcttgcgaga tgatcccgca ttttcttgaa agctttgcag aggctagcag aattaccctc
     5041 cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc gttcaaggct
     5101 cttgcggttg ccataagaga agccacctcg cccaatggta ccaacgatgt tccctccacc
     5161 aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagcatacga tatatataca
     5221 tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt atgatactga
     5281 agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc gctttccttt
     5341 tttctttttg ctttttcttt ttttttctct tgaactcgac ggatctatgc ggtgtgaaat
     5401 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     5461 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5521 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5581 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5641 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5701 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5761 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5821 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5881 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     5941 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     6001 gggtaacgcc agggttttcc cagtcacgac gtt
//
