LOCUS       sequence-239101-        4627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL under PLtetO-1, output promoter PdcuS77.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4627)
  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 4627)
  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: pSR353
FEATURES             Location/Qualifiers
     source          1..4627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(151..739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(231..250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(827..921)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(942..1601)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1516..1535
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1602..1704)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1770..1788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1809..1827,1828..1833,1834..1850,1851..1856,
                     1857..1882)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1809..1827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1834..1852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2588..2659
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2675..2702
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2903..2914
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2921..3634
                     /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      3661..3732
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3748..3775
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3827..3861
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             3896..4522
                     /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(3972..3991)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      4528..4614
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctaggcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
       61 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
      361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
      601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      781 gaacgaaaac tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa
      841 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
      901 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
      961 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacagacg
     1021 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     1081 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     1141 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     1201 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     1261 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     1321 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     1381 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     1441 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     1501 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     1561 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     1621 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     1681 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agatatcgac gtctaagaaa
     1741 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     1801 acctcgagtc cctatcagtg atagagattg acatccctat cagtgataga gatactgagc
     1861 acatcagcag gacgcactga cctaaggatt ttttctagaa gaattatata caacaaatga
     1921 gtaatcagga accggctact atcctgctga ttgacgatca cccgatgctg cgaactggcg
     1981 taaaacagct tatcagtatg gcaccagata tcaccgtggt tggcgaagcg agtaatggcg
     2041 aacagggtat tgaactggcg gagtctcttg atcccgatct gatcctgtta gatctcaata
     2101 tgcccggcat gaacggtctg gaaacgctgg ataaactgcg cgaaaagtcc ctctcagggc
     2161 gcattgtggt attcagcgtc tctaaccatg aagaagatgt ggtcaccgca ctgaaacgcg
     2221 gcgcggatgg ctatctgtta aaagatatgg aaccggaaga tctgctgaaa gcattgcatc
     2281 aggcagctgc tggcgaaatg gtattaagcg aagcattaac gcctgttctg gccgccagct
     2341 tgcgcgctaa ccgtgccact actgagcgcg atgttaacca gttaacccca cgcgagcgcg
     2401 atattctcaa gctgattgcc cagggtttgc cgaacaagat gattgcccgc cgcctggata
     2461 tcaccgaaag cacagtaaaa gtgcacgtca agcacatgct gaagaaaatg aagctcaagt
     2521 ctcgcgtgga agcagcggta tgggtgcatc aggagcgcat tttctgataa taatctagac
     2581 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2641 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     2701 cgtttatagt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa
     2761 tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag ctggttagct
     2821 agtcaagagt ctcttcccct atatagccaa tcattcttac tccctttgaa ttacccgcct
     2881 catcagagat aatgcttaag aaaaagagga gaaatactag atgcgtaaag gcgaagagct
     2941 gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt
     3001 ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat
     3061 ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttatgg
     3121 tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc
     3181 catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa
     3241 aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg
     3301 cattgacttt aaagaagacg gcaatatcct gggccataag ctggaataca attttaacag
     3361 ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat
     3421 tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc
     3481 aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct
     3541 gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc
     3601 gggcatcacg catggtatgg atgaactgta caaatgatga taataatcta gaccaggcat
     3661 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     3721 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     3781 aggtacccgt ggatcctcta gagtcgacct gcaggcatgc aagcttttta cggctagctc
     3841 agtcctaggt atagtgctag cccagccaga gaaacactct ttaacagggg gtagtatgat
     3901 gtctcgttta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     3961 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     4021 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     4081 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     4141 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     4201 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     4261 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgcagtgg ggcattttac
     4321 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     4381 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     4441 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     4501 taaatgtgaa agtgggtctt aaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     4561 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     4621 ccctaga
//
