LOCUS       sequence-247829-        4790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR(1-137aa)-yxdJ(132-229aa) chimera under PLtetO-1, 
            output Promoter PyxdL181.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4790)
  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 4790)
  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: pSR241
FEATURES             Location/Qualifiers
     source          1..4790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..45
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(66..84,85..90,91..107,108..113,114..139)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    66..84
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    91..109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      903..974
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      990..1017
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1322..1333
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1340..2053
                     /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      2080..2151
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2167..2194
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2246..2280
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2315..2941
                     /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(2391..2410)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2947..3033
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3198..3786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3278..3297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3874..3968)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3989..4648)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4563..4582
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcacc
       61 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
      121 tcagcaggac gcactgaccg cataaaggac ttagcaaagc ccaattttaa acaaatgcaa
      181 gagaactaca agattctggt ggtcgatgac gacatgcgcc tgcgtgcgct gctggaacgt
      241 tatctcaccg aacaaggctt ccaggttcga agcgtcgcta atgcagaaca gatggatcgc
      301 ctgctgactc gtgaatcttt ccatcttatg gtactggatt taatgttacc tggtgaagat
      361 ggcttgtcga tttgccgacg tcttcgtagt cagagcaacc cgatgccgat cattatggtg
      421 acggcgaaag gggaagaagt ggaccgtatc gtaggcctgg agattggcgc tgacgactac
      481 attccaaaac cgtttaaccc gcgtgaactg ctggcccgta tccgtgcggt gctgcgtcgt
      541 caggcgaacg aactgccagg cgcaccgtca caggaagagg cggtagttga atatgccggc
      601 gttcagctct ttgtggaacg gtttgaactg cgttttcagg atgaaaaaag tgagctttct
      661 aaaaaagaaa gcaagctttt ggaagtgctg cttgagcggg gagaaaaggt gacgagtcgg
      721 gaccgtctca tggaaaagac gtgggacacc gacatattca tcgatgataa tacacttaac
      781 gtgtatatca cgcggctcag aaaaaaactg cgggagctga atgcgcctgt ttctattgaa
      841 gcggtgcggg gcgaaggcta ccagctgagg gcgcagtcat gataataatc tagaccaggc
      901 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      961 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     1021 atagtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt
     1081 tttgcgccga catcataacg gttctggcaa atattctgaa atgagctggt tagctagtca
     1141 ggacatcagt tcgattttca tttcttacaa aaatgtaagg ctgcgttaag aaaacaacat
     1201 agcccgcttt gtttcccctt gataacatgg atttatgtca aggggatttt tatgttgaac
     1261 gtcccggcac tacatgaaag cagcccagat caacaagaat aaaaccataa ggaggatttt
     1321 caaagaggag aaatactaga tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat
     1381 tctggtggaa ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga
     1441 aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc
     1501 ggtaccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta
     1561 tccggaccat atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca
     1621 ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt
     1681 tgaaggcgat accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg
     1741 caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc
     1801 cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg
     1861 cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct
     1921 gctgccagac aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa
     1981 acgcgatcat atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga
     2041 tgaactgtac aaatgatgat aataatctag accaggcatc aaataaaacg aaaggctcag
     2101 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     2161 cacactggct caccttcggg tgggcctttc tgcgtttata ggtacccgtg gatcctctag
     2221 agtcgacctg caggcatgca agctttttac ggctagctca gtcctaggta tagtgctagc
     2281 ccagccagag aaacactctt taacaggggg tagtatgatg tctcgtttag ataaaagtaa
     2341 agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt taacaacccg
     2401 taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg taaaaaataa
     2461 gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata ctcacttttg
     2521 ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa gttttagatg
     2581 tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc ctacagaaaa
     2641 acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt tttcactaga
     2701 gaatgcatta tatgcactca gcgcagtggg gcattttact ttaggttgcg tattggaaga
     2761 tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata gtatgccgcc
     2821 attattacga caagctatcg aattatttga tcaccaaggt gcagagccag ccttcttatt
     2881 cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa gtgggtctta
     2941 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3001 ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta gggcgttcgg
     3061 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3121 gataacgcat gcaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3601 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3841 cgttaaggga ttttggtcat gactagtgct tggattctca ccaataaaaa acgcccggcg
     3901 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     3961 caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact
     4021 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
     4081 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
     4141 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     4201 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     4261 tttcaccata acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     4321 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     4381 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga aattccggat
     4441 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     4501 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     4561 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     4621 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     4681 actcaaaaaa tacgaccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     4741 cgggccggcc aacgtctcat tttcgccaga tatcgacgtc taagaaacca
//
