LOCUS       sequence-239132-        4674 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of narL(1-126aa)-ydfI(121-213aa) chimera under PLtetO-1, 
            output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4674)
  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 4674)
  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: pSR357.6
FEATURES             Location/Qualifiers
     source          1..4674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(151..739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(231..250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(827..921)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(942..1601)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1516..1535
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1602..1704)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1770..1788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1809..1827,1828..1833,1834..1850,1851..1856,
                     1857..1882)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1809..1827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1834..1852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2597..2668
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2684..2711
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2950..2961
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2968..3681
                     /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      3708..3779
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3795..3822
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3874..3908
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             3943..4569
                     /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(4019..4038)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      4575..4661
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctaggcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
       61 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
      361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
      601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      781 gaacgaaaac tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa
      841 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
      901 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
      961 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacagacg
     1021 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     1081 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     1141 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     1201 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     1261 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     1321 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     1381 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     1441 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     1501 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     1561 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     1621 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     1681 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agatatcgac gtctaagaaa
     1741 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     1801 acctcgagtc cctatcagtg atagagattg acatccctat cagtgataga gatactgagc
     1861 acatcagcag gacgcactga cctaaggatt ttttctagaa gaattatata caacaaatga
     1921 gtaatcagga accggctact atcctgctga ttgacgatca cccgatgctg cgaactggcg
     1981 taaaacagct tatcagtatg gcaccagata tcaccgtggt tggcgaagcg agtaatggcg
     2041 aacagggtat tgaactggcg gagtctcttg atcccgatct gatcctgtta gatctcaata
     2101 tgcccggcat gaacggtctg gaaacgctgg ataaactgcg cgaaaagtcc ctctcagggc
     2161 gcattgtggt attcagcgtc tctaaccatg aagaagatgt ggtcaccgca ctgaaacgcg
     2221 gcgcggatgg ctatctgtta aaagatatgg aaccggaaga tctgctgaaa gcattgcatc
     2281 aggcagctgc tggcaatgtg ctattgcagc ctgatatcct taaacgtctg caagaaatcc
     2341 aatttgagcg gatgaaaaag cagcgcaatg agacgcagct gacagaaaag gaagtcattg
     2401 ttctaaaagc aattgctaaa ggtcttaaaa gcaaagcgat tgcctttgat ttgggcgtct
     2461 ctgagcgaac agtaaagtcc agattaacgt ccatttacaa taaattaggc gcgaattcaa
     2521 gaactgaggc agtaacgatt gccatgcaaa aaggtattct gacaatagac aactaataat
     2581 aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2641 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     2701 gcctttctgc gtttatagta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     2761 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     2821 tggttagcta gtcaactgca tatttgaaaa ttgcccaaac gtacatgccc gaatgtacgt
     2881 ttttttcatt tcattgtcaa ctacaatgag aaagaatgtg atcaagcaat gtgttgaaag
     2941 gagattatca aagaggagaa atactagatg cgtaaaggcg aagagctgtt cactggtgtc
     3001 gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc
     3061 gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt
     3121 aaactgccgg taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt
     3181 gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc
     3241 tatgtgcagg aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa
     3301 gtgaaatttg aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa
     3361 gaagacggca atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac
     3421 atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg
     3481 gaggatggca gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt
     3541 cctgttctgc tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg
     3601 aacgagaaac gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat
     3661 ggtatggatg aactgtacaa atgatgataa taatctagac caggcatcaa ataaaacgaa
     3721 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     3781 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagg tacccgtgga
     3841 tcctctagag tcgacctgca ggcatgcaag ctttttacgg ctagctcagt cctaggtata
     3901 gtgctagccc agccagagaa acactcttta acagggggta gtatgatgtc tcgtttagat
     3961 aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta
     4021 acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta
     4081 aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact
     4141 cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt
     4201 tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct
     4261 acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt
     4321 tcactagaga atgcattata tgcactcagc gcagtggggc attttacttt aggttgcgta
     4381 ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt
     4441 atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc
     4501 ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt
     4561 gggtcttaag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     4621 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc taga
//
