LOCUS       sequence-247895-        4728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of torR(1-134aa)-psdR(134-237aa), ompR137 chimera under 
            PLtetO-1, output Promoter PpsdA110.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4728)
  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 4728)
  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: pKD265
FEATURES             Location/Qualifiers
     source          1..4728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..26
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(47..65,66..71,72..88,89..94,95..120)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    47..65
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    72..90
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      893..964
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      980..1007
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1241..1252
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1259..1972
                     /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      1999..2070
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2086..2113
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2165..2199
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2234..2860
                     /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(2310..2329)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2866..2952
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3117..3705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3197..3216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3793..3887)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3908..4567)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4482..4501
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagtccc tatcagtgat
       61 agagattgac atccctatca gtgatagaga tactgagcac atcagcagga cgcactgacc
      121 taagcgtaaa ataagtaaat tacacctata caagtatgcc acatcacatt gttattgttg
      181 aagatgagcc ggttacccag gcgcgattac aatcctactt cactcaggag gggtataccg
      241 tttccgttac ggcgagcggt gctgggctac gggaaattat gcagaatcag tcggtagatt
      301 taattctgct ggatatcaac ttacccgatg aaaatggcct gatgttaacc cgcgccctgc
      361 gagaacgctc aacggtgggg attattctgg ttaccggacg cagcgatcgg attgaccgta
      421 ttgttgggct ggaaatgggc gcagacgatt acgtcaccaa accgctggaa ctgcgcgaac
      481 tggtagtacg ggtgaaaaat ctgctctggc gaatcgacct cgcgcgacaa gctcaaccgc
      541 acactcagga caactgcgtg gagcttggcg ggctgacgat ttatcctgat cagaatgagg
      601 ccgaatggaa cagcgtccgg attttgtttt cccaaaaaga attccagctt ctctcgattt
      661 ttgtgaggga gcataaaaaa attgtctctc gtgatgaact attggaggcg ttatgggatg
      721 acgttgattt tgtcgatgac aatacgttaa cggtcaatgt caatcggctg cggagaaaac
      781 tggagaatgc aggactgaca gattgcattt ccacaatcag aggacaaggt tatcaatttc
      841 aagtcaatcg gaaggatgaa gcggaatgtt aataataatc tagaccaggc atcaaataaa
      901 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      961 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atagtaaatc
     1021 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga
     1081 catcataacg gttctggcaa atattctgaa atgagctggt tagctagtca caccctcgtg
     1141 aatgtgacag cattgtaaga ttggggagcg gaattgcaag aaagttcgat gggaggatgc
     1201 tgacttcctt tttataataa agaaaaagga ggagcagaac aaagaggaga aatactagat
     1261 gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga
     1321 tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa
     1381 actgacgctg aagttcatct gtactactgg taaactgccg gtaccttggc cgactctggt
     1441 aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca
     1501 tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa
     1561 ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa
     1621 ccgcattgag ctgaaaggca ttgactttaa agaagacggc aatatcctgg gccataagct
     1681 ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat
     1741 taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca
     1801 ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct
     1861 gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct
     1921 ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca aatgatgata
     1981 ataatctaga ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2041 tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt
     2101 gggcctttct gcgtttatag gtacccgtgg atcctctaga gtcgacctgc aggcatgcaa
     2161 gctttttacg gctagctcag tcctaggtat agtgctagcc cagccagaga aacactcttt
     2221 aacagggggt agtatgatgt ctcgtttaga taaaagtaaa gtgattaaca gcgcattaga
     2281 gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc agaagctagg
     2341 tgtagagcag cctacattgt attggcatgt aaaaaataag cgggctttgc tcgacgcctt
     2401 agccattgag atgttagata ggcaccatac tcacttttgc cctttagaag gggaaagctg
     2461 gcaagatttt ttacgtaata acgctaaaag ttttagatgt gctttactaa gtcatcgcga
     2521 tggagcaaaa gtacatttag gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa
     2581 tcaattagcc tttttatgcc aacaaggttt ttcactagag aatgcattat atgcactcag
     2641 cgcagtgggg cattttactt taggttgcgt attggaagat caagagcatc aagtcgctaa
     2701 agaagaaagg gaaacaccta ctactgatag tatgccgcca ttattacgac aagctatcga
     2761 attatttgat caccaaggtg cagagccagc cttcttattc ggccttgaat tgatcatatg
     2821 cggattagaa aaacaactta aatgtgaaag tgggtcttaa ggcatcaaat aaaacgaaag
     2881 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2941 agtaggacaa atccgccgcc ctagacctag ggcgttcggc tgcggcgagc ggtatcagct
     3001 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcatg caagaacatg
     3061 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3121 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3181 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3241 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3301 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3361 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3421 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3481 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3541 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3601 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3661 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3721 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3781 actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     3841 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
     3901 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     3961 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     4021 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     4081 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     4141 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccataa cacgccacat
     4201 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     4261 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     4321 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     4381 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     4441 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     4501 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     4561 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgaccggta
     4621 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gggccggcca acgtctcatt
     4681 ttcgccagat atcgacgtct aagaaaccat tattatcatg acattaac
//
