LOCUS       sequence-247751-        4672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-126aa)-liaR(122-211aa) chimera under PLtetO-1, 
            output Promoter PliaI121.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4672)
  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 4672)
  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: pSR358.2
FEATURES             Location/Qualifiers
     source          1..4672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..64
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(85..103,104..109,110..126,127..132,133..158)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    85..103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    110..128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      864..935
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      951..978
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1223..1234
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1241..1954
                     /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      1981..2052
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2068..2095
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2147..2181
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2216..2842
                     /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(2292..2311)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2848..2934
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3099..3687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3179..3198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3775..3869)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3890..4549)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4464..4483
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 atcgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
       61 aggccctttc gtcttcacct cgagtcccta tcagtgatag agattgacat ccctatcagt
      121 gatagagata ctgagcacat cagcaggacg cactgaccta aggatttttt ctagaagaat
      181 tatatacaac aaatgagtaa tcaggaaccg gctactatcc tgctgattga cgatcacccg
      241 atgctgcgaa ctggcgtaaa acagcttatc agtatggcac cagatatcac cgtggttggc
      301 gaagcgagta atggcgaaca gggtattgaa ctggcggagt ctcttgatcc cgatctgatc
      361 ctgttagatc tcaatatgcc cggcatgaac ggtctggaaa cgctggataa actgcgcgaa
      421 aagtccctct cagggcgcat tgtggtattc agcgtctcta accatgaaga agatgtggtc
      481 accgcactga aacgcggcgc ggatggctat ctgttaaaag atatggaacc ggaagatctg
      541 ctgaaagcat tgcatcaggc agctgctggc gagccgaagc tggagtcaaa agtggcggga
      601 aaagtattat ccaggctgcg ccactcaggt gaaaacgcgc tcccgcatga atcgcttaca
      661 aaacgggagc tcgaaatact ctgcctgatc gcagaaggaa agacaaacaa agaaataggc
      721 gaggaactgt ttattacgat taaaacagtc aaaacacata ttacgaatat tttatcaaag
      781 ctggatgtca gtgaccggac gcaggcggcg gtgtacgcac accgaaatca tctcgtgaat
      841 tagtaataat ctagaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      901 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
      961 cgggtgggcc tttctgcgtt tatagtaaat cactgcataa ttcgtgtcgc tcaaggcgca
     1021 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
     1081 aatgagctgg ttagctagtc agtgccaatg ggtccggtgc gagatacgac tccggtctta
     1141 tataaaaatc aatctctgat tcgttttgca tatcttccaa cttgtataag atgaagacaa
     1201 ggaaaacgaa aggaggatct gcaaagagga gaaatactag atgcgtaaag gcgaagagct
     1261 gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt
     1321 ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat
     1381 ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttatgg
     1441 tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc
     1501 catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa
     1561 aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg
     1621 cattgacttt aaagaagacg gcaatatcct gggccataag ctggaataca attttaacag
     1681 ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat
     1741 tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc
     1801 aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct
     1861 gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc
     1921 gggcatcacg catggtatgg atgaactgta caaatgatga taataatcta gaccaggcat
     1981 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2041 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     2101 aggtacccgt ggatcctcta gagtcgacct gcaggcatgc aagcttttta cggctagctc
     2161 agtcctaggt atagtgctag cccagccaga gaaacactct ttaacagggg gtagtatgat
     2221 gtctcgttta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     2281 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     2341 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     2401 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     2461 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     2521 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     2581 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgcagtgg ggcattttac
     2641 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     2701 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     2761 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     2821 taaatgtgaa agtgggtctt aaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     2881 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     2941 ccctagacct agggcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3001 cggttatcca cagaatcagg ggataacgca tgcaagaaca tgtgagcaaa aggccagcaa
     3061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgactagtgc ttggattctc
     3781 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     3841 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     3901 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     3961 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     4021 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     4081 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     4141 ctttagggaa ataggccagg ttttcaccat aacacgccac atcttgcgaa tatatgtgta
     4201 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     4261 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     4321 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     4381 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     4441 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     4501 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     4561 tagctcctga aaatctcgat aactcaaaaa atacgaccgg tagtgatctt atttcattat
     4621 ggtgaaagtt ggaacctctt acgggccggc caacgtctca ttttcgccag at
//
