LOCUS       sequence-245242-        4705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4487(1-127aa)-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 4705)
  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 4705)
  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: pKD283.2
FEATURES             Location/Qualifiers
     source          1..4705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..82
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(103..121,122..127,128..144,145..150,151..176)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    103..121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    128..146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      926..997
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1013..1040
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1274..1285
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1292..2005
                     /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      2032..2103
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2119..2146
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2198..2232
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2267..2893
                     /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(2343..2362)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2899..2985
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3150..3738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3230..3249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3826..3920)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3941..4600)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4515..4534
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtctcatttt cgccagatat cgacgtctaa gaaaccatta ttatcatgac attaacctat
       61 aaaaataggc gtatcacgag gccctttcgt cttcacctcg agtccctatc agtgatagag
      121 attgacatcc ctatcagtga tagagatact gagcacatca gcaggacgca ctgaccaatc
      181 cattaaacca ataaaaaccc tcagtttaca tgcgcttact gctgattgaa gatgatatcg
      241 acttagtggc tcgccttatt cccgcactta acaaagcggg ctatacggtt gaacacgccg
      301 ataacggcat cgatggcgcc tttctgggcg aggaagaagc atttgaggct gtgatcctcg
      361 acttaggatt gcccggtaag ccgggcttaa gcgtgctaag tcactggcgt caaaaaggct
      421 tggctatgcc cgtccttatc cttaccgccc gcgatgcgtg gcacgaacgc gttgatggcc
      481 tcaaggccgg cgccgatgat tatcttggca agccttttca tgttgaagag ttacttgcca
      541 gattagaagc gctgatccgc cgccactttg gccgtgccga taacctactg gtggagcttg
      601 gcgggctgac gatttatcct gatcagaatg aggccgaatg gaacagcgtc cggattttgt
      661 tttcccaaaa agaattccag cttctctcga tttttgtgag ggagcataaa aaaattgtct
      721 ctcgtgatga actattggag gcgttatggg atgacgttga ttttgtcgat gacaatacgt
      781 taacggtcaa tgtcaatcgg ctgcggagaa aactggagaa tgcaggactg acagattgca
      841 tttccacaat cagaggacaa ggttatcaat ttcaagtcaa tcggaaggat gaagcggaat
      901 gttaataata atctagacca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
      961 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     1021 ttcgggtggg cctttctgcg tttatagtaa atcactgcat aattcgtgtc gctcaaggcg
     1081 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     1141 gaaatgagct ggttagctag tcacaccctc gtgaatgtga cagcattgta agattgggga
     1201 gcggaattgc aagaaagttc gatgggagga tgctgacttc ctttttataa taaagaaaaa
     1261 ggaggagcag aacaaagagg agaaatacta gatgcgtaaa ggcgaagagc tgttcactgg
     1321 tgtcgtccct attctggtgg aactggatgg tgatgtcaac ggtcataagt tttccgtgcg
     1381 tggcgagggt gaaggtgacg caactaatgg taaactgacg ctgaagttca tctgtactac
     1441 tggtaaactg ccggtacctt ggccgactct ggtaacgacg ctgacttatg gtgttcagtg
     1501 ctttgctcgt tatccggacc atatgaagca gcatgacttc ttcaagtccg ccatgccgga
     1561 aggctatgtg caggaacgca cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc
     1621 ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt gagctgaaag gcattgactt
     1681 taaagaagac ggcaatatcc tgggccataa gctggaatac aattttaaca gccacaatgt
     1741 ttacatcacc gccgataaac aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa
     1801 cgtggaggat ggcagcgtgc agctggctga tcactaccag caaaacactc caatcggtga
     1861 tggtcctgtt ctgctgccag acaatcacta tctgagcacg caaagcgttc tgtctaaaga
     1921 tccgaacgag aaacgcgatc atatggttct gctggagttc gtaaccgcag cgggcatcac
     1981 gcatggtatg gatgaactgt acaaatgatg ataataatct agaccaggca tcaaataaaa
     2041 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2101 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggtacccg
     2161 tggatcctct agagtcgacc tgcaggcatg caagcttttt acggctagct cagtcctagg
     2221 tatagtgcta gcccagccag agaaacactc tttaacaggg ggtagtatga tgtctcgttt
     2281 agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg
     2341 tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca
     2401 tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca
     2461 tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa
     2521 aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg
     2581 gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg
     2641 tttttcacta gagaatgcat tatatgcact cagcgcagtg gggcatttta ctttaggttg
     2701 cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga
     2761 tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc
     2821 agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga
     2881 aagtgggtct taaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2941 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc
     3001 tagggcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3061 acagaatcag gggataacgc atgcaagaac atgtgagcaa aaggccagca aaaggccagg
     3121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3781 aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct caccaataaa
     3841 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     3901 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
     3961 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     4021 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     4081 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     4141 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     4201 aataggccag gttttcacca taacacgcca catcttgcga atatatgtgt agaaactgcc
     4261 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     4321 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     4381 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     4441 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     4501 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     4561 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     4621 aaaatctcga taactcaaaa aatacgaccg gtagtgatct tatttcatta tggtgaaagt
     4681 tggaacctct tacgggccgg ccaac
//
