LOCUS       sequence-243342-        6033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN HIS3 CNB1.
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: pTJK502
FEATURES             Location/Qualifiers
     source          1..6033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(816..832)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(817..833)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(861..879)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(900..916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(900..916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(913..935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    924..940
                     /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(948..954,955..972,973..978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    993..1014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1131..1148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1302..1890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1382..1401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2061..2852,2853..2921))
                     /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     2684..2703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2922..3026)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3056..3559
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3607..3625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3663..3685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3785..3804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3951..4138
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             4139..4801
                     /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(5061..5516)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5198..5219)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5410..5429
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5642..5664
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5656..5673
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5657..5673
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5680..5699
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5680..5698
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5731..5747
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5731..5747
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gtaactcaat ggtgatcaga atccatagaa gcatttttat ttcttaaaat gggtgctgct
       61 ccttccaaaa ttgtggatgg tcttttagaa gatacaaatt gtatgtacac ttcggagtga
      121 ggaaaagaaa gaaaggggaa attaaccgat tttactaaca ctgacacttt gaacagttga
      181 tagagatgaa attgaaaggt taaggaagag attcatgaaa ttagatagag atagctcagg
      241 gtctattgat aaaaatgaat ttatgagcat tcctggcgtt tcgtcaaacc ctcttgctgg
      301 acgtataatg gaggttttcg atgctgataa tagtggggac gtggattttc aagagttcat
      361 cacaggatta tccattttca gtgggcgtgg gtccaaggac gaaaagttaa gattcgcctt
      421 caaaatctac gacattgaca aggacggttt catatccaat ggtgagttgt tcatcgtgtt
      481 gaagattatg gtaggttcta atctggacga tgaacagctg caacagatag tagataggac
      541 gatagtggaa aacgatagcg acggcgacgg acgtttaagt ttcgaggagt ttaagaatgc
      601 tatcgaaacc acagaagtgg ccaagagtct gacattgcaa tacgatgtgt aagactaggg
      661 gacacttcat tcatttatgg tatgccaata tttttaagaa aagaagaata atacgcgata
      721 ttgtttttta aggaaggaac gcacactcgc ccagttagag tgctgatgat atatacatat
      781 atatatgtat atgtaacaaa caataagaag cttatcgata ccgtcgacct cgaggggggg
      841 cccggtacca gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg
      901 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      961 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1021 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1081 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1861 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1921 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1981 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2041 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2101 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2161 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2221 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2281 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2341 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2401 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2461 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2521 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2581 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2641 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2701 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2761 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2821 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2881 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2941 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3001 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacggatcg
     3061 cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac
     3121 tctgtgttta tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta
     3181 gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag
     3241 cgtactttac atatatattt attagacaag aaaagcagat taaatagata tacattcgat
     3301 taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag
     3361 atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg
     3421 ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat
     3481 ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt aaattataat
     3541 tatttttata gcacgtgatg aaaaggaccc taagaaacca ttattatcat gacattaacc
     3601 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3661 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3721 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3781 tatgcggcat cagagcagat tgtactgaga gtgcaccata aattcccgtt ttaagagctt
     3841 ggtgagcgct aggagtcact gccaggtatc gtttgaacac ggcattagtc agggaagtca
     3901 taacacagtc ctttcccgca attttctttt tctattactc ttggcctcct ctagtacact
     3961 ctatattttt ttatgcctcg gtaatgattt tcattttttt ttttccacct agcggatgac
     4021 tctttttttt tcttagcgat tggcattatc acataatgaa ttatacatta tataaagtaa
     4081 tgtgatttct tcgaagaata tactaaaaaa tgagcaggca agataaacga aggcaaagat
     4141 gacagagcag aaagccctag taaagcgtat tacaaatgaa accaagattc agattgcgat
     4201 ctctttaaag ggtggtcccc tagcgataga gcactcgatc ttcccagaaa aagaggcaga
     4261 agcagtagca gaacaggcca cacaatcgca agtgattaac gtccacacag gtatagggtt
     4321 tctggaccat atgatacatg ctctggccaa gcattccggc tggtcgctaa tcgttgagtg
     4381 cattggtgac ttacacatag acgaccatca caccactgaa gactgcggga ttgctctcgg
     4441 tcaagctttt aaagaggccc taggggccgt gcgtggagta aaaaggtttg gatcaggatt
     4501 tgcgcctttg gatgaggcac tttccagagc ggtggtagat ctttcgaaca ggccgtacgc
     4561 agttgtcgaa cttggtttgc aaagggagaa agtaggagat ctctcttgcg agatgatccc
     4621 gcattttctt gaaagctttg cagaggctag cagaattacc ctccacgttg attgtctgcg
     4681 aggcaagaat gatcatcacc gtagtgagag tgcgttcaag gctcttgcgg ttgccataag
     4741 agaagccacc tcgcccaatg gtaccaacga tgttccctcc accaaaggtg ttcttatgta
     4801 gtgacaccga ttatttaaag ctgcagcata cgatatatat acatgtgtat atatgtatac
     4861 ctatgaatgt cagtaagtat gtatacgaac agtatgatac tgaagatgac aaggtaatgc
     4921 atcattctat acgtgtcatt ctgaacgagg cgcgctttcc ttttttcttt ttgctttttc
     4981 tttttttttc tcttgaactc gacggatcta tgcggtgtga aataccgcac agatgcgtaa
     5041 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     5101 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5161 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     5221 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     5281 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     5341 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     5401 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     5461 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     5521 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5581 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     5641 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     5701 gaattggagc tccaccgcgg tggcggccgc tctagaacta gtggatccac gctatgaaaa
     5761 aggaaaacaa atgactctta taatttgttt ccctcgactt ctctatttaa atccaagatt
     5821 ttaactaata aactatttaa taataaaaca attaactctt tatatggaag ccttggaaat
     5881 tagccgccaa aatgggatat acattccgtg cgaagtgacc gcgtggaagg ccgggtatca
     5941 tcttaaaaat cactagtttc ttttttagcg gaatgcaata aaggtgcttt gtgctggtgt
     6001 ggtttacacg gaacatctag tagctaaaac ttg
//
