LOCUS       sequence-313208-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert: Pfus1mut-tetR-nls-malE-tCyc1. Same insert as pCL33-decoder 
            in a multiple integration shuttle vector. Backbone: pRG235 
            (addgene). Marker: Leu2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  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 7025)
  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: pCL66-decoder
FEATURES             Location/Qualifiers
     source          1..7025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(111..699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(191..210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(878..1669,1670..1738))
                     /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     1501..1520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1739..1843)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(2155..3249)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFQNHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     3327..3349
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3341..3358
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3342..3358
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3365..3384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3365..3383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4220..4239)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             4765..4785
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6134..6150)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6135..6151)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      6178..6427
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6194..6212)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(6659..6677)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6698..6714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6698..6714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6711..6733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6722..6738
                     /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(6965..6982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
       61 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      121 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      181 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      241 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      301 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      361 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      421 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      481 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      541 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      601 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      661 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttgt
      721 ttaaacctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      781 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
      841 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
      901 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      961 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1021 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1081 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1141 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1201 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1261 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1321 tggttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1381 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1441 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1501 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1561 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1621 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1681 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     1741 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     1801 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     1861 tgccacctga cgtcgaaaga acgcggcatt agcaacattt gcgaagcgaa cgattttgcg
     1921 aactgctatt aattaactct ggcgcgccac tatcgtctat caactaatag ttatattatc
     1981 aatatattat catatacggt gttaagatga tgacataagt tatgagaagc tgtcatcgaa
     2041 gttagaggaa gctgaagtgc aaggattgat aatgtaatag gataatcata tgtttctatt
     2101 atgaatttca tttataaagt ttatgtacaa atatcataaa aaaagagaat ctttttaagc
     2161 aaggattttc ttaacttctt cggcgacagc atcaccgact tcggtggtac tgttggaacc
     2221 acctaaatca ccagttctga tacctgcatc caaaaccttt ttaactgcat cttcaatggc
     2281 cttaccttct tcaggcaagt tcaatgacaa tttcaacatc attgcagcag acaagatagt
     2341 ggcgataggg ttgaccttat tctttggcaa atctggagca gaaccgtggc atggttcgta
     2401 caaaccaaat gcggtgttct tgtctggcaa agaggccaag gacgcagatg gcaacaaacc
     2461 caaggaacct gggataacgg aggcttcatc ggagatgatg tcaccaaaca tgttgctggt
     2521 gattataata ccatttaggt gggttgggtt cttaactagg atcatggcgg cagaatcaat
     2581 caattgatgt tgaaccttca atgtaggaaa ttcgttcttg atggtttcct ccacagtttt
     2641 tctccataat cttgaagagg ccaaaacatt agctttatcc aaggaccaaa taggcaatgg
     2701 tggctcatgt tgtagggcca tgaaagcggc cattcttgtg attctttgca cttctggaac
     2761 ggtgtattgt tcactatccc aagcgacacc atcaccatcg tcttcctttc tcttaccaaa
     2821 gtaaatacct cccactaatt ctctgacaac aacgaagtca gtacctttag caaattgtgg
     2881 cttgattgga gataagtcta aaagagagtc ggatgcaaag ttacatggtc ttaagttggc
     2941 gtacaattga agttctttac ggatttttag taaaccttgt tcaggtctaa cactaccagt
     3001 accccattta ggaccaccca cagcacctaa caaaacggca tcagccttct tggaggcttc
     3061 cagcgcctca tctggaagtg gaacacctgt agcatcgata gcagcaccac caattaaatg
     3121 attttggaaa tcgaacttga cattggaacg aacatcagaa atagctttaa gaaccttaat
     3181 ggcttcggct gtgatttctt gaccaacgtg gtcacctggc aaaacgacga tcttcttagg
     3241 ggcagacatt agaatggtat atccttgaaa tatatatacg atcggatgtg ctgcaaggcg
     3301 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3361 attgtaatac gactcactat agggcgaatt ggagaaacag aataatatat aaacctgtat
     3421 aatataacct tgaagactat atttcttttc gcggccgcct atagggcgaa ttggagaaac
     3481 agaataatat ataaacctgt ataatataac cttgaagact atatttcttt tcgcggccgc
     3541 ctatagggcg aattggagct ccctaggacg atgattcagt tcgccttcta tcctttgttt
     3601 acgtatttgt ttatatatat aactttattt ttttttatta attgggctgc aagacaattt
     3661 tgttgtcagt gatgcctcaa tccttctttt gcttccatat ttaccatgtg gaccctttca
     3721 aaacagagtt gtatctctgc aggatgccct ttttgacgta ttgaatggca taattgcact
     3781 gtcacttttc gcgctgtctc attttggtgc gatgatgaaa caaacatgaa acgtctgtaa
     3841 tttgaaacaa ataacgtaat tctcgggatt ggttttattt aaatgacaat gtaagagtgg
     3901 ctttgtaagg tatgtgttgc tctagaaaag gttgaaacca gttccctgaa attattcccc
     3961 tactattcat tgaatagtat aaaagctatt cattgaatag gtaattctgt aaatctattt
     4021 cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta aaacaccaag
     4081 aacttagttt cgaataaaca cacataaaca aacaaaacta gtggatcccc cgggctgcag
     4141 gaattcatgt ccagattaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat
     4201 gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag
     4261 cctacattgt attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag
     4321 atgttagata ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt
     4381 ttacgtaata acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa
     4441 gtacatttag gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc
     4501 tttttatgcc aacaaggttt ttcactagag aatgcattat atgcactcag cgctgtgggg
     4561 cattttactt taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg
     4621 gaaacaccta ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat
     4681 caccaaggtg cagagccagc cttcttattc ggccttgaat tgatcatttg cggattagaa
     4741 aaacaactta aatgtgaaag tgcccccaag aaaaagagaa aggtggcaga agccgccgca
     4801 aaggaagcgg cagcaaaaga ggctgcggcc aaggaagctg ctgccaaggc gctagaagca
     4861 gaggcggccg caaaggaggc ggccgcgaag gaagcggctg cgaaggaggc agcggcaaag
     4921 gcgaaaataa agaccggggc gaggattctt gccttgtccg cgttaacaac aatgatgttt
     4981 tctgcaagcg ccctagcaaa aatcgaggag ggaaagttag ttatatggat caatggtgac
     5041 aaaggataca atgggctagc tgaagtcggg aaaaagtttg aaaaggacac ggggataaaa
     5101 gtcacggtgg agcatccgga caaactagaa gagaaatttc ctcaggtagc cgcaaccggg
     5161 gatgggcccg acattatatt ttgggcacat gatcgtttcg ggggatacgc ccaaagcggt
     5221 cttcttgctg aaataactcc tgataaagca ttccaagata agctttaccc tttcacatgg
     5281 gacgccgtta ggtataacgg caaactgatt gcttatccta tagctgtgga ggcactatct
     5341 ttgatttaca acaaagacct tttgccaaac cctcccaaga cttgggaaga gataccagcc
     5401 ctagataaag aattaaaagc taagggaaag tccgcgctaa tgttcaatct tcaagaacca
     5461 tattttactt ggccgttaat agctgctgat ggtggatatg cctttaagta cgagaacggg
     5521 aagtacgaca ttaaagatgt tggcgttgac aatgccggag cgaaggctgg tttaactttc
     5581 cttgtggact taatcaaaaa taagcacatg aacgcggata cagactattc catcgccgaa
     5641 gctgctttca ataagggaga gaccgctatg acgatcaacg ggccatgggc atggtcaaat
     5701 attgatacca gtaaagtaaa ttatggagta accgttcttc cgaccttcaa aggacagcct
     5761 tctaagccct tcgtgggggt cctttcagcg ggcatcaacg cagcaagccc taataaggaa
     5821 ttggcgaaag agtttcttga gaactatcta ctaactgacg agggtttaga ggctgtaaac
     5881 aaagacaagc ctcttggcgc agtcgccctt aaatcttacg aggaagaact agctaaagat
     5941 ccgaggatag ccgctaccat ggagaatgct caaaaaggag aaatcatgcc aaacatcccg
     6001 caaatgtctg cattttggta tgctgtacgt acggcagtga ttaacgccgc ttctggaaga
     6061 cagaccgttg acgaagcgct taaagacgcc cagtaaactt ctaaataagc gaatttacgc
     6121 gtttaaagct tatcgatacc gtcgacctcg agacaggccc cttttccttt gtctatatca
     6181 tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa
     6241 aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt
     6301 agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca aacgcgtgta
     6361 cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt
     6421 aatttgcaag gttcgcagtt tacactctca tcgtcgctct catcatcgct tccgttgttg
     6481 ttttccttag tagcgtctgc ttggtaccca gcttcggccg tatttacaga taacgatgtc
     6541 gttattatat atatatatat atattgtcac ttgtacccag cttcggccgt atttacagat
     6601 aacgatgtcg ttattatata tatatatata tattgtcact tgtacccagc ttttgttccc
     6661 tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6721 attgttatcc gctcacaaca gctggaaaca tataaaacgg aatgaggaat aatcgtaata
     6781 ttagtatgta gaaatataga ttccattttg aggattccta tatccttgag gagaacttct
     6841 agtatattct gtatacctaa tattatagcc tttatcaaca atggcgcgcc tctccctgca
     6901 ggcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7021 actca
//
