LOCUS       sequence-238763-        7435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CEN TRP1 CNA1-Delta-AID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7435)
  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 7435)
  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: pTJK518
FEATURES             Location/Qualifiers
     source          1..7435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1777..1795)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1816..1832)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1816..1832)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1829..1851)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1840..1856
                     /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(1864..1870,1871..1888,1889..1894))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1909..1930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2047..2064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2218..2806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2298..2317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2977..3768,3769..3837))
                     /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     3600..3619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3838..3942)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3979..4482
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4523..4541
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4579..4601)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4701..4720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4741..5021
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5022..5696
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(5799..6254)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5936..5957)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6148..6167
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6380..6402
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6394..6411
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6395..6411
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6418..6437
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6418..6436
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     6469..6485
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6469..6485
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 atgtcgaaag acttgaattc ttcacgcctc aaaatcatta aacctaatga ctcttacata
       61 aaggttgacc ggaaaaaaga tttgacaaaa tacgaattag aaaacggtaa agtaatttct
      121 actaaggacc gacctatcgc ttctgtacct gccataacag gaaagatacc aagtgatgag
      181 gaagtattcg actccaagac gggattacct aatcattcct ttttaagaga gcatttcttt
      241 catgagggtc gactttctaa ggaacaggcc ataaaaatct taaatatgtc aactgtagca
      301 ttgagtaaag aacccaatct actaaaactc aaagcgccaa ttactatatg tggtgatatt
      361 cacgggcagt attatgattt attgaaactg tttgaagttg gcggtgaccc cgccgaaatc
      421 gactatttat tcttggggga ttatgttgat agaggtgcat tctcttttga gtgtctgatt
      481 tatttgtact ccttgaagtt gaataattta gggagatttt ggatgctaag aggtaaccat
      541 gagtgtaagc acttgacctc ttattttact tttaagaatg agatgttgca caaatacgat
      601 atggaagttt acgatgcttg ctgcagatca ttcaatgtct taccattagc agctttaatg
      661 aacggacaat atttttgtgt gcatggtggt atctctccag agttaaaatc agtagaggat
      721 gttaataaaa ttaatagatt tcgagaaatc ccatctcgtg gtctcatgtg tgacctacta
      781 tgggccgatc ctgtcgaaaa ttatgatgat gcaagagatg gtagcgaatt tgatcagagc
      841 gaggatgaat tcgtacctaa cagtttgagg ggttgctctt tcgccttcac ttttaaagca
      901 tcatgcaagt ttttgaaggc aaatggtttg ttatctatta ttagagcaca cgaagcacag
      961 gatgctgggt acagaatgta taaaaacaat aaagtaacag gcttcccgag cttaataacc
     1021 atgttcagtg cgccaaacta cctggacaca tatcataata aagctgctgt gttaaaatat
     1081 gaagaaaacg tcatgaacat caggcagttt cacatgtctc cgcaccctta ctggttgcct
     1141 gattttatgg atgttttcac ctggtcacta ccttttgttg gcgaaaaagt tactagcatg
     1201 ttagtgtcta tattaaacat atgtagtgag caggaacttg acccagaatc ggaacccaaa
     1261 gctgcggagg agactgtaaa ggcaagagca aacgcaacta aggagaccgg caccccatct
     1321 gatgaaaagg cgtcatcagc gatattagaa gatgaaaccc aactgtgata aatctcattt
     1381 tattcttttt tattcctata agatgcaaca tcgaattctt gtgctactgt cacgatccgc
     1441 aagagataat atgaactaac aaaatacgaa gaaaatatag atcatttcca gtctatttct
     1501 tgatttccat acttaactgc gagcagaata aatataaaca catatttttt agttttattt
     1561 tattattaat gaaccctagt taccaattag ttatgtaaac gcttttcgta agaatgtaag
     1621 ttatgaacac caccttctag ttgagcagaa gcggttctaa attcaaatct aactttaccg
     1681 ctttggacat tttcctttaa taaagttagg gacttgtaac caatgctcga atcttgctga
     1741 aaaactcgag ggggggcccg gtaccagctt ttgttccctt tagtgagggt taatttcgag
     1801 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1861 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1921 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1981 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2041 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2101 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2161 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2221 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2281 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2341 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2401 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2461 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2521 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2581 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2641 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2701 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2761 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2821 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2881 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2941 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3001 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3061 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3121 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3181 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3241 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3301 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3361 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3421 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3481 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3541 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3601 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3661 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3721 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3781 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3841 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3901 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3961 gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     4021 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     4081 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     4141 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     4201 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     4261 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     4321 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     4381 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     4441 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtctaag aaaccattat
     4501 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4561 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4621 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4681 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagatc
     4741 acgacattac tatatatata atataggaag catttaatag aacagcatcg taatatatgt
     4801 gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag
     4861 cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta
     4921 attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca
     4981 cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca
     5041 caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat
     5101 gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa
     5161 caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt
     5221 caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg
     5281 ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag
     5341 aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca
     5401 acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag
     5461 aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc
     5521 aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg
     5581 gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa
     5641 atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta
     5701 ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa
     5761 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     5821 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5881 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5941 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6001 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6061 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6121 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6181 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6241 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6301 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6361 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
     6421 tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc tagaactagt
     6481 ggatccgtct tcgtcaacaa ttggttcagg tttagacgat actaagtcag gtacaacctc
     6541 gtccatctct tcatcttcat cttcgtcatc atagtcctct tcatcttcat cgtcttcatc
     6601 atcatcaccc tcaatatgca caactctttt tgatttttta gttctctgct tttcttgcca
     6661 ttccaaatac aatttttcta ccttattata atttaaattg aaacagtcgt tatatatgtt
     6721 gatgacctct acggcaatcg gtaaagccga gtcgtcttca acattagttt caaattcatc
     6781 tatcattgca taaagtaacg tttcttcgat caaagcggcg tcgacaactt tttcattttt
     6841 gaaaaggtct actacaatcc ctgtaatcca gtctctcttc tcagctgagt ctggaccacc
     6901 ccaactgttt tctacggcaa catccaacgc atcccactta taaataacca tggaaacacc
     6961 gagctcaaaa cgtgcctgtt gcttttcgtc agagaacatt agattggttt tgccttgctc
     7021 agcctgaaca aatgctgtct catcaatata ttgtgtgctc attgttccgt aaggattatc
     7081 actataaaaa ttttccgcag atttcttttg gactccacct ttcttcctgt tgtaataaat
     7141 acaaactatt tgaatcaatt tgatattcat ctatatctat ttcaaaattt ttcatgtcat
     7201 cgcctcttga aacgtgaatt ttccaattct gaaaaagaac gtactactgg gaaacaaaag
     7261 ggaaaaatgt ataaatcctt taatgttttt aaatcaagag gcattattat aaaagaacga
     7321 agcaaagcct ttaatatttg ctttattaaa ggtactattc aaagaaaagt ttttttagat
     7381 tctttttttt tttttcacgt attagctcag ctgccataaa acactctcaa cgcca
//
