LOCUS       sequence-313201-        6753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert: Pfus1mut-tetR-nls-malE-tCyc1. TetR-NLS-MBP (Azizoglu et al, 
            bioRxiv 2019) is inducible by alpha-factor. Backbone: pRG203MX 
            (addgene). Marker: His3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6753)
  AUTHORS   Lormeau C, Rudolf F, Stelling J
  TITLE     A rationally engineered decoder of transient intracellular signals.
  JOURNAL   Nat Commun. 2021 Mar 25;12(1):1886. doi: 10.1038/s41467-021-22190-4.
  PUBMED    33767179
REFERENCE   2  (bases 1 to 6753)
  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"; journalName: "Nat 
            Commun."; date: "2021-03-25"; volume: "12(1):1886. doi"; pages: " 
            10.1038/s41467-021-22190-4"
COMMENT     Sequence Label: pCL33-decoder
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     199..221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     213..230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     214..230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     237..256
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        237..255
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1010..1029)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1555..1575
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2924..2940)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2925..2941)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2968..3217
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2984..3002)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3449..3467)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3488..3504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3488..3504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3501..3523)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3512..3528
                     /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)."
     primer_bind     complement(3762..3779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3933..4521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4013..4032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4700..5491,5492..5560))
                     /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     5323..5342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5561..5665)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5963..6613
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQK
                     GEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQA
                     MGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLYS
                     FSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
ORIGIN
        1 tacgtagccg tataaattga attgaatgtt cagcttatag aatagagaca ctttgctgtt
       61 caatgcgtcg tcacttacca tactcacttt attatacgac tttaagtata aactccgcgg
      121 ttatggtaaa attaatgatg cacaaacgtc cgatcggatg tgctgcaagg cgattaagtt
      181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
      241 acgactcact atagggcgaa ttggagaaac agaataatat ataaacctgt ataatataac
      301 cttgaagact atatttcttt tcgcggccgc ctatagggcg aattggagct ccctaggacg
      361 atgattcagt tcgccttcta tcctttgttt acgtatttgt ttatatatat aactttattt
      421 ttttttatta attgggctgc aagacaattt tgttgtcagt gatgcctcaa tccttctttt
      481 gcttccatat ttaccatgtg gaccctttca aaacagagtt gtatctctgc aggatgccct
      541 ttttgacgta ttgaatggca taattgcact gtcacttttc gcgctgtctc attttggtgc
      601 gatgatgaaa caaacatgaa acgtctgtaa tttgaaacaa ataacgtaat tctcgggatt
      661 ggttttattt aaatgacaat gtaagagtgg ctttgtaagg tatgtgttgc tctagaaaag
      721 gttgaaacca gttccctgaa attattcccc tactattcat tgaatagtat aaaagctatt
      781 cattgaatag gtaattctgt aaatctattt cttaaacttc ttaaattcta cttttatagt
      841 tagtcttttt tttagtttta aaacaccaag aacttagttt cgaataaaca cacataaaca
      901 aacaaaacta gtggatcccc cgggctgcag gaattcatgt ccagattaga taaaagtaaa
      961 gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt
     1021 aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag
     1081 cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc
     1141 cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt
     1201 gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa
     1261 cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag
     1321 aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt attggaagat
     1381 caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca
     1441 ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc
     1501 ggccttgaat tgatcatttg cggattagaa aaacaactta aatgtgaaag tgcccccaag
     1561 aaaaagagaa aggtggcaga agccgccgca aaggaagcgg cagcaaaaga ggctgcggcc
     1621 aaggaagctg ctgccaaggc gctagaagca gaggcggccg caaaggaggc ggccgcgaag
     1681 gaagcggctg cgaaggaggc agcggcaaag gcgaaaataa agaccggggc gaggattctt
     1741 gccttgtccg cgttaacaac aatgatgttt tctgcaagcg ccctagcaaa aatcgaggag
     1801 ggaaagttag ttatatggat caatggtgac aaaggataca atgggctagc tgaagtcggg
     1861 aaaaagtttg aaaaggacac ggggataaaa gtcacggtgg agcatccgga caaactagaa
     1921 gagaaatttc ctcaggtagc cgcaaccggg gatgggcccg acattatatt ttgggcacat
     1981 gatcgtttcg ggggatacgc ccaaagcggt cttcttgctg aaataactcc tgataaagca
     2041 ttccaagata agctttaccc tttcacatgg gacgccgtta ggtataacgg caaactgatt
     2101 gcttatccta tagctgtgga ggcactatct ttgatttaca acaaagacct tttgccaaac
     2161 cctcccaaga cttgggaaga gataccagcc ctagataaag aattaaaagc taagggaaag
     2221 tccgcgctaa tgttcaatct tcaagaacca tattttactt ggccgttaat agctgctgat
     2281 ggtggatatg cctttaagta cgagaacggg aagtacgaca ttaaagatgt tggcgttgac
     2341 aatgccggag cgaaggctgg tttaactttc cttgtggact taatcaaaaa taagcacatg
     2401 aacgcggata cagactattc catcgccgaa gctgctttca ataagggaga gaccgctatg
     2461 acgatcaacg ggccatgggc atggtcaaat attgatacca gtaaagtaaa ttatggagta
     2521 accgttcttc cgaccttcaa aggacagcct tctaagccct tcgtgggggt cctttcagcg
     2581 ggcatcaacg cagcaagccc taataaggaa ttggcgaaag agtttcttga gaactatcta
     2641 ctaactgacg agggtttaga ggctgtaaac aaagacaagc ctcttggcgc agtcgccctt
     2701 aaatcttacg aggaagaact agctaaagat ccgaggatag ccgctaccat ggagaatgct
     2761 caaaaaggag aaatcatgcc aaacatcccg caaatgtctg cattttggta tgctgtacgt
     2821 acggcagtga ttaacgccgc ttctggaaga cagaccgttg acgaagcgct taaagacgcc
     2881 cagtaaactt ctaaataagc gaatttacgc gtttaaagct tatcgatacc gtcgacctcg
     2941 agacaggccc cttttccttt gtctatatca tgtaattagt tatgtcacgc ttacattcac
     3001 gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     3061 tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat
     3121 ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac attatactga aaaccttgct
     3181 tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag gttcgcagtt tacactctca
     3241 tcgtcgctct catcatcgct tccgttgttg ttttccttag tagcgtctgc ttggtaccca
     3301 gcttcggccg tatttacaga taacgatgtc gttattatat atatatatat atattgtcac
     3361 ttgtacccag cttcggccgt atttacagat aacgatgtcg ttattatata tatatatata
     3421 tattgtcact tgtacccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     3481 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaaca gctgacaccg
     3541 attatttaaa gctgcagcat acgatatata tacatgtgta tatatgtata cctatgaatg
     3601 tcagtaagta tgtatacgaa cagtatgata ctgaagatga caaggtaatg catcattcta
     3661 tacgtgtcat tctgaacgag gcgcgcctct ccctgcaggc attaatgaat cggccaacgc
     3721 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3781 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4501 cgcagaaaaa aaggatctca agaagatcct ttgatctttt gtttaaacct acggggtctg
     4561 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4621 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4801 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4861 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4921 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4981 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5041 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5101 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatggttgtc agaagtaagt
     5161 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5221 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5281 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5341 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5401 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5461 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5521 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5581 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5641 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgaaa
     5701 gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta ttaattaact
     5761 ctggcgcgcc tcctctagta cactctatat ttttttatgc ctcggtaatg attttcattt
     5821 ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat tatcacataa
     5881 tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa aaaatgagca
     5941 ggcaagataa acgaaggcac atatgaggag ggcttttgta gaaagaaata cgaacgaaac
     6001 gaaaatcagc gttgccatcg ctttggacaa agctccctta cctgaagagt cgaattttat
     6061 tgatgaactt ataacttcca agcatgcaaa ccaaaaggga gaacaagtaa tccaagtaga
     6121 cacgggaatt ggattcttgg atcacatgta tcatgcactg gctaaacatg caggctggag
     6181 cttacgactt tactcaagag gtgatttaat catcgatgat catcacactg cagaagatac
     6241 tgctattgca cttggtattg cattcaagca ggctatgggt aactttgccg gtgttaaaag
     6301 atttggacat gcttattgtc cacttgacga agctctttcc agaagcgtag ttgacttgtc
     6361 gggacggccc tatgctgtta tcgatttggg attaaagcgt gaaaaggttg gggaattgtc
     6421 ctgtgaaatg atccctcact tactatattc cttttcggta gctgctggaa ttactttgca
     6481 tgttacctgc ttatatggta gtaatgacca tcatcgtgct gaaagcgctt ttaaatctct
     6541 ggctgttgcc atgcgcgcgg ctacttctct tactggaagt tctgaagtcc caagcacgaa
     6601 gggagtgttg taaagatgaa ttggattatg tcaggaaaag aacgacaatt ttgcatccaa
     6661 attgtctaaa ttttagagtt gcttgaaaac aatagaacct tacttgcttt ataattacgt
     6721 taattagaag cgttatctcg tgaaggaata tag
//
