LOCUS       sequence-247793-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-159aa)-liaR(154-211aa) chimera under PLtetO-1, 
            output Promoter PliaI121.
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: pSR358.4
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..54
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(75..93,94..99,100..116,117..122,123..148)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    75..93
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    100..118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      857..928
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      944..971
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1216..1227
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1234..1947
                     /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      1974..2045
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2061..2088
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2140..2174
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2209..2835
                     /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(2285..2304)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2841..2927
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3092..3680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3172..3191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3768..3862)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3883..4542)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4457..4476
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
       61 gtcttcacct cgagtcccta tcagtgatag agattgacat ccctatcagt gatagagata
      121 ctgagcacat cagcaggacg cactgaccta aggatttttt ctagaagaat tatatacaac
      181 aaatgagtaa tcaggaaccg gctactatcc tgctgattga cgatcacccg atgctgcgaa
      241 ctggcgtaaa acagcttatc agtatggcac cagatatcac cgtggttggc gaagcgagta
      301 atggcgaaca gggtattgaa ctggcggagt ctcttgatcc cgatctgatc ctgttagatc
      361 tcaatatgcc cggcatgaac ggtctggaaa cgctggataa actgcgcgaa aagtccctct
      421 cagggcgcat tgtggtattc agcgtctcta accatgaaga agatgtggtc accgcactga
      481 aacgcggcgc ggatggctat ctgttaaaag atatggaacc ggaagatctg ctgaaagcat
      541 tgcatcaggc agctgctggc gaaatggtat taagcgaagc attaacgcct gttctggccg
      601 ccagcttgcg cgctaaccgt gccactactg agcgcgatgt taaccagtta accccacgcg
      661 agctcgaaat actctgcctg atcgcagaag gaaagacaaa caaagaaata ggcgaggaac
      721 tgtttattac gattaaaaca gtcaaaacac atattacgaa tattttatca aagctggatg
      781 tcagtgaccg gacgcaggcg gcggtgtacg cacaccgaaa tcatctcgtg aattagtaat
      841 aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
      901 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
      961 gcctttctgc gtttatagta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     1021 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     1081 tggttagcta gtcagtgcca atgggtccgg tgcgagatac gactccggtc ttatataaaa
     1141 atcaatctct gattcgtttt gcatatcttc caacttgtat aagatgaaga caaggaaaac
     1201 gaaaggagga tctgcaaaga ggagaaatac tagatgcgta aaggcgaaga gctgttcact
     1261 ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg
     1321 cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact
     1381 actggtaaac tgccggtacc ttggccgact ctggtaacga cgctgactta tggtgttcag
     1441 tgctttgctc gttatccgga ccatatgaag cagcatgact tcttcaagtc cgccatgccg
     1501 gaaggctatg tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt
     1561 gcggaagtga aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac
     1621 tttaaagaag acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat
     1681 gtttacatca ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac
     1741 aacgtggagg atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt
     1801 gatggtcctg ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa
     1861 gatccgaacg agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc
     1921 acgcatggta tggatgaact gtacaaatga tgataataat ctagaccagg catcaaataa
     1981 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2041 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tataggtacc
     2101 cgtggatcct ctagagtcga cctgcaggca tgcaagcttt ttacggctag ctcagtccta
     2161 ggtatagtgc tagcccagcc agagaaacac tctttaacag ggggtagtat gatgtctcgt
     2221 ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt cggaatcgaa
     2281 ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac attgtattgg
     2341 catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt agataggcac
     2401 catactcact tttgcccttt agaaggggaa agctggcaag attttttacg taataacgct
     2461 aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca tttaggtaca
     2521 cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt atgccaacaa
     2581 ggtttttcac tagagaatgc attatatgca ctcagcgcag tggggcattt tactttaggt
     2641 tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac acctactact
     2701 gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca aggtgcagag
     2761 ccagccttct tattcggcct tgaattgatc atatgcggat tagaaaaaca acttaaatgt
     2821 gaaagtgggt cttaaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     2881 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgccctaga
     2941 cctagggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3001 ccacagaatc aggggataac gcatgcaaga acatgtgagc aaaaggccag caaaaggcca
     3061 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3121 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3181 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3241 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3301 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3361 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3421 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3481 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3541 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3601 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3721 ggaacgaaaa ctcacgttaa gggattttgg tcatgactag tgcttggatt ctcaccaata
     3781 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     3841 actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca
     3901 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     3961 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     4021 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     4081 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     4141 gaaataggcc aggttttcac cataacacgc cacatcttgc gaatatatgt gtagaaactg
     4201 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     4261 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     4321 acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     4381 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     4441 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     4501 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     4561 tgaaaatctc gataactcaa aaaatacgac cggtagtgat cttatttcat tatggtgaaa
     4621 gttggaacct cttacgggcc ggccaacgtc tcattttcgc cagatatcga cgtct
//
