LOCUS       sequence-245251-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-129aa)-ydfI(124-213aa) chimera under PLtetO-1, 
            output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4675)
  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 4675)
  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: pSR357.9
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      780..851
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      867..894
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1133..1144
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1151..1864
                     /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      1891..1962
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1978..2005
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2057..2091
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2126..2752
                     /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(2202..2221)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2758..2844
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3009..3597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3089..3108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3685..3779)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3800..4459)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4374..4393
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4628..4646
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4667..4675,1..10,11..16,17..33,34..39,40..65)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    join(4667..4675,1..10)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gtgatagaga ttgacatccc tatcagtgat agagatactg agcacatcag caggacgcac
       61 tgacctaagg attttttcta gaagaattat atacaacaaa tgagtaatca ggaaccggct
      121 actatcctgc tgattgacga tcacccgatg ctgcgaactg gcgtaaaaca gcttatcagt
      181 atggcaccag atatcaccgt ggttggcgaa gcgagtaatg gcgaacaggg tattgaactg
      241 gcggagtctc ttgatcccga tctgatcctg ttagatctca atatgcccgg catgaacggt
      301 ctggaaacgc tggataaact gcgcgaaaag tccctctcag ggcgcattgt ggtattcagc
      361 gtctctaacc atgaagaaga tgtggtcacc gcactgaaac gcggcgcgga tggctatctg
      421 ttaaaagata tggaaccgga agatctgctg aaagcattgc atcaggcagc tgctggcgaa
      481 atggtattgc agcctgatat ccttaaacgt ctgcaagaaa tccaatttga gcggatgaaa
      541 aagcagcgca atgagacgca gctgacagaa aaggaagtca ttgttctaaa agcaattgct
      601 aaaggtctta aaagcaaagc gattgccttt gatttgggcg tctctgagcg aacagtaaag
      661 tccagattaa cgtccattta caataaatta ggcgcgaatt caagaactga ggcagtaacg
      721 attgccatgc aaaaaggtat tctgacaata gacaactaat aataatctag accaggcatc
      781 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
      841 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
      901 gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt
      961 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctggttag ctagtcaact
     1021 gcatatttga aaattgccca aacgtacatg cccgaatgta cgtttttttc atttcattgt
     1081 caactacaat gagaaagaat gtgatcaagc aatgtgttga aaggagatta tcaaagagga
     1141 gaaatactag atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga
     1201 actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc
     1261 aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg
     1321 gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca
     1381 tatgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac
     1441 gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga
     1501 taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct
     1561 gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca
     1621 aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca
     1681 gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga
     1741 caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca
     1801 tatggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta
     1861 caaatgatga taataatcta gaccaggcat caaataaaac gaaaggctca gtcgaaagac
     1921 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     1981 tcaccttcgg gtgggccttt ctgcgtttat aggtacccgt ggatcctcta gagtcgacct
     2041 gcaggcatgc aagcttttta cggctagctc agtcctaggt atagtgctag cccagccaga
     2101 gaaacactct ttaacagggg gtagtatgat gtctcgttta gataaaagta aagtgattaa
     2161 cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc
     2221 ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt
     2281 gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt gccctttaga
     2341 aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact
     2401 aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga
     2461 aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt
     2521 atatgcactc agcgcagtgg ggcattttac tttaggttgc gtattggaag atcaagagca
     2581 tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg
     2641 acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga
     2701 attgatcata tgcggattag aaaaacaact taaatgtgaa agtgggtctt aaggcatcaa
     2761 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2821 aacgctctcc tgagtaggac aaatccgccg ccctagacct agggcgttcg gctgcggcga
     2881 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2941 tgcaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3001 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3061 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3121 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3181 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3241 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3301 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3361 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3421 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3481 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3541 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3601 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3661 attttggtca tgactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     3721 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     3781 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     3841 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     3901 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     3961 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     4021 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccat
     4081 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     4141 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     4201 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     4261 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     4321 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     4381 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     4441 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     4501 atacgaccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgggccggc
     4561 caacgtctca ttttcgccag atatcgacgt ctaagaaacc attattatca tgacattaac
     4621 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagtccc tatca
//
