LOCUS       sequence-251167-        4644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-159aa)-uhpA(142-129aa) chimera under PLtetO-1, 
            output Promoter PuhpT99.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4644)
  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 4644)
  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: pSR359.4
FEATURES             Location/Qualifiers
     source          1..4644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..73
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(94..112,113..118,119..135,136..141,142..167)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    94..112
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    119..137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      867..938
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      954..981
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1204..1215
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1222..1935
                     /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      1962..2033
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2049..2076
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2128..2162
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2197..2823
                     /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(2273..2292)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2829..2915
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3080..3668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3160..3179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3756..3850)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3871..4530)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4445..4464
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcgccagata tcgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
       61 cgtatcacga ggccctttcg tcttcacctc gagtccctat cagtgataga gattgacatc
      121 cctatcagtg atagagatac tgagcacatc agcaggacgc actgacctaa ggattttttc
      181 tagaagaatt atatacaaca aatgagtaat caggaaccgg ctactatcct gctgattgac
      241 gatcacccga tgctgcgaac tggcgtaaaa cagcttatca gtatggcacc agatatcacc
      301 gtggttggcg aagcgagtaa tggcgaacag ggtattgaac tggcggagtc tcttgatccc
      361 gatctgatcc tgttagatct caatatgccc ggcatgaacg gtctggaaac gctggataaa
      421 ctgcgcgaaa agtccctctc agggcgcatt gtggtattca gcgtctctaa ccatgaagaa
      481 gatgtggtca ccgcactgaa acgcggcgcg gatggctatc tgttaaaaga tatggaaccg
      541 gaagatctgc tgaaagcatt gcatcaggca gctgctggcg aaatggtatt aagcgaagca
      601 ttaacgcctg ttctggccgc cagcttgcgc gctaaccgtg ccactactga gcgcgatgtt
      661 aaccagttaa ccccacgcga acgccaggtg gcggaaaaac tggcgcaagg aatggcggtg
      721 aaagagattg ccgccgaact gggcttgtca ccgaaaacgg tacacgtcca tcgcgccaat
      781 ctgatggaaa aactgggcgt cagtaacgac gtagagctgg cgcgccgcat gtttgatggc
      841 tggtgataat aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
      961 cttcgggtgg gcctttctgc gtttatagta aatcactgca taattcgtgt cgctcaaggc
     1021 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
     1081 tgaaatgagc tggttagcta gtcatttcac ttcatatccg gcaaaactaa gaaattttcc
     1141 aggttttgcc tggacgctat ctcaggcctg atttgctgct gatttttaca atgcatgcct
     1201 cacaaagagg agaaatacta gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct
     1261 attctggtgg aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt
     1321 gaaggtgacg caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg
     1381 ccggtacctt ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt
     1441 tatccggacc atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg
     1501 caggaacgca cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa
     1561 tttgaaggcg ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagac
     1621 ggcaatatcc tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc
     1681 gccgataaac aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat
     1741 ggcagcgtgc agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt
     1801 ctgctgccag acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag
     1861 aaacgcgatc atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg
     1921 gatgaactgt acaaatgatg ataataatct agaccaggca tcaaataaaa cgaaaggctc
     1981 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     2041 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggtacccg tggatcctct
     2101 agagtcgacc tgcaggcatg caagcttttt acggctagct cagtcctagg tatagtgcta
     2161 gcccagccag agaaacactc tttaacaggg ggtagtatga tgtctcgttt agataaaagt
     2221 aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc
     2281 cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat
     2341 aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt
     2401 tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga
     2461 tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa
     2521 aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta
     2581 gagaatgcat tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa
     2641 gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg
     2701 ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta
     2761 ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct
     2821 taaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     2881 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc tagggcgttc
     2941 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3001 gggataacgc atgcaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3481 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3721 cacgttaagg gattttggtc atgactagtg cttggattct caccaataaa aaacgcccgg
     3781 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
     3841 aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta
     3901 ctgttgtaat tcattaagca ttcttccgac atggaagcca tcacaaacgg catgatgaac
     3961 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     4021 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     4081 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     4141 gttttcacca taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     4201 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     4261 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg
     4321 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     4381 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     4441 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     4501 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     4561 taactcaaaa aatacgaccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     4621 tacgggccgg ccaacgtctc attt
//
