LOCUS       sequence-247802-        4628 bp    DNA     circular SYN 15-APR-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 4628)
  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 4628)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..4628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..19
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(40..58,59..64,65..81,82..87,88..113)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    40..58
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    65..83
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      819..890
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      906..933
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1134..1145
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1152..1865
                     /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      1892..1963
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1979..2006
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2058..2092
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2127..2753
                     /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(2203..2222)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2759..2845
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3010..3598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3090..3109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3686..3780)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3801..4460)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4375..4394
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aataggcgta tcacgaggcc ctttcgtctt cacctcgagt ccctatcagt gatagagatt
       61 gacatcccta tcagtgatag agatactgag cacatcagca ggacgcactg acctaaggat
      121 tttttctaga agaattatat acaacaaatg agtaatcagg aaccggctac tatcctgctg
      181 attgacgatc acccgatgct gcgaactggc gtaaaacagc ttatcagtat ggcaccagat
      241 atcaccgtgg ttggcgaagc gagtaatggc gaacagggta ttgaactggc ggagtctctt
      301 gatcccgatc tgatcctgtt agatctcaat atgcccggca tgaacggtct ggaaacgctg
      361 gataaactgc gcgaaaagtc cctctcaggg cgcattgtgg tattcagcgt ctctaaccat
      421 gaagaagatg tggtcaccgc actgaaacgc ggcgcggatg gctatctgtt aaaagatatg
      481 gaaccggaag atctgctgaa agcattgcat caggcagctg ctggcgaaat ggtattaagc
      541 gaagcattaa cgcctgttct ggccgccagc ttgcgcgcta accgtgccac tactgagcgc
      601 gatgttaacc agttaacccc acgcgagcgc gatattctca agctgattgc ccagggtttg
      661 ccgaacaaga tgattgcccg ccgcctggat atcaccgaaa gcacagtaaa agtgcacgtc
      721 aagcacatgc tgaagaaaat gaagctcaag tctcgcgtgg aagcagcggt atgggtgcat
      781 caggagcgca ttttctgata ataatctaga ccaggcatca aataaaacga aaggctcagt
      841 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
      901 acactggctc accttcgggt gggcctttct gcgtttatag taaatcactg cataattcgt
      961 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     1021 tggcaaatat tctgaaatga gctggttagc tagtcaagag tctcttcccc tatatagcca
     1081 atcattctta ctccctttga attacccgcc tcatcagaga taatgcttaa gaaaaagagg
     1141 agaaatacta gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg
     1201 aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg
     1261 caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt
     1321 ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc
     1381 atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca
     1441 cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg
     1501 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc
     1561 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac
     1621 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc
     1681 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag
     1741 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc
     1801 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt
     1861 acaaatgatg ataataatct agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1921 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     1981 ctcaccttcg ggtgggcctt tctgcgttta taggtacccg tggatcctct agagtcgacc
     2041 tgcaggcatg caagcttttt acggctagct cagtcctagg tatagtgcta gcccagccag
     2101 agaaacactc tttaacaggg ggtagtatga tgtctcgttt agataaaagt aaagtgatta
     2161 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     2221 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     2281 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     2341 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     2401 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     2461 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     2521 tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     2581 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     2641 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     2701 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaggcatca
     2761 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2821 gaacgctctc ctgagtagga caaatccgcc gccctagacc tagggcgttc ggctgcggcg
     2881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2941 atgcaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3421 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3661 gattttggtc atgactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga
     3721 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     3781 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     3841 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     3901 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     3961 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     4021 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcacca
     4081 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     4141 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     4201 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
     4261 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     4321 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     4381 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     4441 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     4501 aatacgaccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgggccgg
     4561 ccaacgtctc attttcgcca gatatcgacg tctaagaaac cattattatc atgacattaa
     4621 cctataaa
//
