LOCUS       sequence-245432-        4813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ompR under PLtetO-1, output promoter PompF146.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4813)
  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 4813)
  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: pSR62
FEATURES             Location/Qualifiers
     source          1..4813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(95..113,114..119,120..136,137..142,143..168)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    95..113
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    120..138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      944..1015
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1031..1058
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1200..1243)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             1392..2105
                     /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      2132..2203
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2219..2246
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2298..2332
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2367..2993
                     /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(2443..2462)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2999..3085
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3250..3838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3330..3349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3926..4020)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4041..4700)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4615..4634
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttcgccagat atcgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
       61 gcgtatcacg aggccctttc gtcttcacct cgagtcccta tcagtgatag agattgacat
      121 ccctatcagt gatagagata ctgagcacat cagcaggacg cactgaccgc ataaaggact
      181 tagcaaagcc caattttaaa caaatgcaag agaactacaa gattctggtg gtcgatgacg
      241 acatgcgcct gcgtgcgctg ctggaacgtt atctcaccga acaaggcttc caggttcgaa
      301 gcgtcgctaa tgcagaacag atggatcgcc tgctgactcg tgaatctttc catcttatgg
      361 tactggattt aatgttacct ggtgaagatg gcttgtcgat ttgccgacgt cttcgtagtc
      421 agagcaaccc gatgccgatc attatggtga cggcgaaagg ggaagaagtg gaccgtatcg
      481 taggcctgga gattggcgct gacgactaca ttccaaaacc gtttaacccg cgtgaactgc
      541 tggcccgtat ccgtgcggtg ctgcgtcgtc aggcgaacga actgccaggc gcaccgtcac
      601 aggaagaggc ggtaattgct ttcggtaagt tcaaacttaa cctcggtacg cgcgaaatgt
      661 tccgcgaaga cgagccgatg ccgctcacca gcggtgagtt tgcggtactg aaggcactgg
      721 tcagccatcc gcgtgagccg ctctcccgcg ataagctgat gaaccttgcc cgtggtcgtg
      781 aatattccgc aatggaacgc tccatcgacg tgcagatttc gcgtctgcgc cgcatggtgg
      841 aagaagatcc agcgcatccg cgttacattc agaccgtctg gggtctgggc tacgtctttg
      901 taccggacgg ctctaaagca tgataataat ctagaccagg catcaaataa aacgaaaggc
      961 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     1021 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagtaaat cactgcataa
     1081 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     1141 ggttctggca aatattctga aatgagctgg ttagctagtc aggcgtatca cgaggcagaa
     1201 tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggttcc ttaaatttta
     1261 cttttggtta catatttttt ctttttgaaa ccaaatcttt atctttgtag cactttcacg
     1321 gtagcgaaac gttagtttga atggaaagat gcctgcaggt gtcataaaaa aaaccatgag
     1381 ggtaataaat aatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg
     1441 aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg
     1501 caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt
     1561 ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc
     1621 atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca
     1681 cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg
     1741 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc
     1801 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac
     1861 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc
     1921 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag
     1981 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc
     2041 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt
     2101 acaaatgatg ataataatct agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2161 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     2221 ctcaccttcg ggtgggcctt tctgcgttta taggtacccg tggatcctct agagtcgacc
     2281 tgcaggcatg caagcttttt acggctagct cagtcctagg tatagtgcta gcccagccag
     2341 agaaacactc tttaacaggg ggtagtatga tgtctcgttt agataaaagt aaagtgatta
     2401 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     2461 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     2521 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     2581 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     2641 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     2701 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     2761 tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     2821 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     2881 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     2941 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaggcatca
     3001 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3061 gaacgctctc ctgagtagga caaatccgcc gccctagacc tagggcgttc ggctgcggcg
     3121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3181 atgcaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3661 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3901 gattttggtc atgactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga
     3961 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     4021 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     4081 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     4141 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     4201 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     4261 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcacca
     4321 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     4381 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     4441 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
     4501 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     4561 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     4621 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     4681 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     4741 aatacgaccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgggccgg
     4801 ccaacgtctc att
//
