LOCUS       sequence-238771-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2µm HIS3 CNB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7759)
  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 7759)
  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: pTJK507
FEATURES             Location/Qualifiers
     source          1..7759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..70)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     261..280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     493..515
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     507..524
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     508..524
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     531..550
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        531..549
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     582..598
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     582..598
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1700..1716)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1701..1717)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1745..1763)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1784..1800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1784..1800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1797..1819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1808..1824
                     /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(1832..1838,1839..1856,1857..1862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1877..1898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2015..2032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2186..2774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2266..2285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2945..3736,3737..3805))
                     /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     3568..3587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3806..3910)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4495..5837
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5458..5505)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     6217..6235
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6273..6295)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6395..6414
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6561..6748
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             6749..7411
                     /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(join(7671..7759,1..367))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
       61 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
      121 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
      181 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
      241 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
      301 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
      361 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      421 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      481 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
      541 cactataggg cgaattggag ctccaccgcg gtggcggccg ctctagaact agtggatcca
      601 cgctatgaaa aaggaaaaca aatgactctt ataatttgtt tccctcgact tctctattta
      661 aatccaagat tttaactaat aaactattta ataataaaac aattaactct ttatatggaa
      721 gccttggaaa ttagccgcca aaatgggata tacattccgt gcgaagtgac cgcgtggaag
      781 gccgggtatc atcttaaaaa tcactagttt cttttttagc ggaatgcaat aaaggtgctt
      841 tgtgctggtg tggtttacac ggaacatcta gtagctaaaa cttggtaact caatggtgat
      901 cagaatccat agaagcattt ttatttctta aaatgggtgc tgctccttcc aaaattgtgg
      961 atggtctttt agaagataca aattgtatgt acacttcgga gtgaggaaaa gaaagaaagg
     1021 ggaaattaac cgattttact aacactgaca ctttgaacag ttgatagaga tgaaattgaa
     1081 aggttaagga agagattcat gaaattagat agagatagct cagggtctat tgataaaaat
     1141 gaatttatga gcattcctgg cgtttcgtca aaccctcttg ctggacgtat aatggaggtt
     1201 ttcgatgctg ataatagtgg ggacgtggat tttcaagagt tcatcacagg attatccatt
     1261 ttcagtgggc gtgggtccaa ggacgaaaag ttaagattcg ccttcaaaat ctacgacatt
     1321 gacaaggacg gtttcatatc caatggtgag ttgttcatcg tgttgaagat tatggtaggt
     1381 tctaatctgg acgatgaaca gctgcaacag atagtagata ggacgatagt ggaaaacgat
     1441 agcgacggcg acggacgttt aagtttcgag gagtttaaga atgctatcga aaccacagaa
     1501 gtggccaaga gtctgacatt gcaatacgat gtgtaagact aggggacact tcattcattt
     1561 atggtatgcc aatattttta agaaaagaag aataatacgc gatattgttt tttaaggaag
     1621 gaacgcacac tcgcccagtt agagtgctga tgatatatac atatatatat gtatatgtaa
     1681 caaacaataa gaagcttatc gataccgtcg acctcgaggg ggggcccggt accagctttt
     1741 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg
     1801 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     1861 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1921 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1981 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2041 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2101 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2761 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2881 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2941 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3001 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3061 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3121 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3181 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3241 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3301 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3361 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3421 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3481 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3541 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3601 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3661 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3721 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3781 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3841 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3901 gggttccgcg cacatttccc cgaaaagtgc cacctaattc cacggactat agactatact
     3961 agtatactcc gtctactgta cgatacactt ccgctcaggt ccttgtcctt taacgaggcc
     4021 ttaccactct tttgttactc tattgatcca gctcagcaaa ggcagtgtga tctaagattc
     4081 tatcttcgcg atgtagtaaa actagctaga ccgagaaaga gactagaaat gcaaaaggca
     4141 cttctacaat ggctgccatc attattatcc gatgtgacgc tgcagcttct caatgatatt
     4201 cgaatacgct ttgaggagat acagcctaat atccgacaaa ctgttttaca gatttacgat
     4261 cgtacttgtt acccatcatt gaattttgaa catccgaacc tgggagtttt ccctgaaaca
     4321 gatagtatat ttgaacctgt ataataatat atagtctagc gctttacgga agacaatgta
     4381 tgtatttcgg ttcctggaga aactattgca tctattgcat aggtaatctt gcacgtcgca
     4441 tccccggttc attttctgcg tttccatctt gcacttcaat agcatatctt tgttaacgaa
     4501 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     4561 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     4621 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     4681 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     4741 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     4801 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     4861 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     4921 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     4981 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     5041 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     5101 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     5161 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     5221 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     5281 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     5341 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     5401 gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa
     5461 gttcctatac tttctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa
     5521 acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca
     5581 cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt
     5641 tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc
     5701 tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt
     5761 agctgttcta tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt
     5821 tcctttgata ttggatcata tgcatagtac cgagaaacta gtgcgaagta gtgatcaggt
     5881 attgctgtta tctgatgagt atacgttgtc ctggccacgg cagaagcacg cttatcgctc
     5941 caatttccca caacattagt caactccgtt aggcccttca ttgaaagaaa tgaggtcatc
     6001 aaatgtcttc caatgtgaga ttttgggcca ttttttatag caaagattga ataaggcgca
     6061 tttttcttca aagctttatt gtacgatctg actaagttat cttttaataa ttggtattcc
     6121 tgtttattgc ttgaagaatt gccggtccta tttactcgtt ttaggactgg ttcagaattc
     6181 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6241 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6301 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6361 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6421 gtgcaccata aattcccgtt ttaagagctt ggtgagcgct aggagtcact gccaggtatc
     6481 gtttgaacac ggcattagtc agggaagtca taacacagtc ctttcccgca attttctttt
     6541 tctattactc ttggcctcct ctagtacact ctatattttt ttatgcctcg gtaatgattt
     6601 tcattttttt ttttccacct agcggatgac tctttttttt tcttagcgat tggcattatc
     6661 acataatgaa ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa
     6721 tgagcaggca agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat
     6781 tacaaatgaa accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga
     6841 gcactcgatc ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca
     6901 agtgattaac gtccacacag gtatagggtt tctggaccat atgatacatg ctctggccaa
     6961 gcattccggc tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca
     7021 caccactgaa gactgcggga ttgctctcgg tcaagctttt aaagaggccc taggggccgt
     7081 gcgtggagta aaaaggtttg gatcaggatt tgcgcctttg gatgaggcac tttccagagc
     7141 ggtggtagat ctttcgaaca ggccgtacgc agttgtcgaa cttggtttgc aaagggagaa
     7201 agtaggagat ctctcttgcg agatgatccc gcattttctt gaaagctttg cagaggctag
     7261 cagaattacc ctccacgttg attgtctgcg aggcaagaat gatcatcacc gtagtgagag
     7321 tgcgttcaag gctcttgcgg ttgccataag agaagccacc tcgcccaatg gtaccaacga
     7381 tgttccctcc accaaaggtg ttcttatgta gtgacaccga ttatttaaag ctgcagcata
     7441 cgatatatat acatgtgtat atatgtatac ctatgaatgt cagtaagtat gtatacgaac
     7501 agtatgatac tgaagatgac aaggtaatgc atcattctat acgtgtcatt ctgaacgagg
     7561 cgcgctttcc ttttttcttt ttgctttttc tttttttttc tcttgaactc gacggatcta
     7621 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     7681 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     7741 ataggccgaa atcggcaaa
//
