LOCUS       sequence-247778-        4677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-154aa)-ydfI(148-213aa) chimera under PLtetO-1, 
            output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4677)
  AUTHORS   Schmidl SR, Ekness F, Sofjan K, Daeffler KN, Brink KR, Landry BP, 
            Gerhardt KP, Dyulgyarov N, Sheth RU, Tabor JJ
  TITLE     Rewiring bacterial two-component systems by modular DNA-binding 
            domain swapping.
  JOURNAL   Nat Chem Biol. 2019 Jul;15(7):690-698. doi: 
            10.1038/s41589-019-0286-6. Epub 2019 May 20.
  PUBMED    31110305
REFERENCE   2  (bases 1 to 4677)
  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: 
            "10.1038/s41589-019-0286-6"; journalName: "Nat Chem Biol"; date: 
            "2019-07"; volume: "15"; issue: "7"; pages: "690-698"
COMMENT     Sequence Label: pSR357.20
FEATURES             Location/Qualifiers
     source          1..4677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..19
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(40..58,59..64,65..81,82..87,88..113)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    40..58
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    65..83
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      831..902
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      918..945
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1183..1194
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1201..1914
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1941..2012
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2028..2055
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2107..2141
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2176..2802
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2252..2271)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2808..2894
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3059..3647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3139..3158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3735..3829)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3850..4509)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4424..4443
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aataggcgta tcacgaggcc ctttcgtctt cacctcgagt ccctatcagt gatagagatt
       61 gacatcccta tcagtgatag agatactgag cacatcagca ggacgcactg acctaaggat
      121 tttttctaga agaattatat acaacaaatg agtaatcagg aaccggctac tatcctgctg
      181 attgacgatc acccgatgct gcgaactggc gtaaaacagc ttatcagtat ggcaccagat
      241 atcaccgtgg ttggcgaagc gagtaatggc gaacagggta ttgaactggc ggagtctctt
      301 gatcccgatc tgatcctgtt agatctcaat atgcccggca tgaacggtct ggaaacgctg
      361 gataaactgc gcgaaaagtc cctctcaggg cgcattgtgg tattcagcgt ctctaaccat
      421 gaagaagatg tggtcaccgc actgaaacgc ggcgcggatg gctatctgtt aaaagatatg
      481 gaaccggaag atctgctgaa agcattgcat caggcagctg ctggcgaaat ggtattaagc
      541 gaagcattaa cgcctgttct ggccgccagc ttgcgcgcta accgtgccac tactgagcgc
      601 gatgttaacc agctgacaga aaaggaagtc attgttctaa aagcaattgc taaaggtctt
      661 aaaagcaaag cgattgcctt tgatttgggc gtctctgagc gaacagtaaa gtccagatta
      721 acgtcaattt acaataaatt aggcgcgaat tcaagaactg aggcagtaac gattgccatg
      781 caaaaaggta ttctgacaat agacaactaa taataatcta gaccaggcat caaataaaac
      841 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
      901 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat agtaaatcac
      961 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
     1021 tcataacggt tctggcaaat attatgaaat gagctggtta gctagtcaac tgcatattta
     1081 aaattgccca aacgtacatg cccgaatgta cgtttttttc atttcattgt caactacaat
     1141 gagaaagaat gtgatcaagc aatgtgttga aaggagatta tcaaagagga gaaatactag
     1201 atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt
     1261 gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt
     1321 aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg gccgactctg
     1381 gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag
     1441 catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt
     1501 aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta
     1561 aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct gggccataag
     1621 ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc
     1681 attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat
     1741 cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat
     1801 ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg
     1861 ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgatga
     1921 taataatcta gaccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1981 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
     2041 gtgggccttt ctgcgtttat aggtacccgt ggatcctcta gagtcgacct gcaggcatgc
     2101 aagcttttta cggctagctc agtcctaggt atagtgctag cccagccaga gaaacactct
     2161 ttaacagggg gtagtatgat gtctcgttta gataaaagta aagtgattaa cagcgcatta
     2221 gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta
     2281 ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc
     2341 ttagccattg agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc
     2401 tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc
     2461 gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa
     2521 aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc
     2581 agcgcagtgg ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct
     2641 aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc
     2701 gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata
     2761 tgcggattag aaaaacaact taaatgtgaa agtgggtctt aaggcatcaa ataaaacgaa
     2821 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     2881 tgagtaggac aaatccgccg ccctagacct agggcgttcg gctgcggcga gcggtatcag
     2941 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca tgcaagaaca
     3001 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3061 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3121 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3181 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3241 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3301 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3361 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3421 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3481 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3541 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3601 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3661 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3721 tgactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     3781 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     3841 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     3901 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     3961 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     4021 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     4081 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccat aacacgccac
     4141 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     4201 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     4261 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc
     4321 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     4381 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     4441 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     4501 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgaccgg
     4561 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgggccggc caacgtctca
     4621 ttttcgccag atatcgacgt ctaagaaacc attattatca tgacattaac ctataaa
//
