LOCUS       sequence-455061-        3826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid expressing NanoLuciferase under the control of the 
            promoter PydfJ115, cognate to YdfI. Responds to NarL/NarX chimeras 
            from this study..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3826)
  AUTHORS   Peruzzi JA, Galvez NR, Kamat NP
  TITLE     Engineering transmembrane signal transduction in synthetic membranes
            using two-component systems.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 May 9;120(19):e2218610120. doi: 
            10.1073/pnas.2218610120. Epub 2023 May 1.
  PUBMED    37126679
REFERENCE   2  (bases 1 to 3826)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2023-05-9"; volume: "120(19):e2218610120. 
            doi"; pages: " 10.1073/pnas.2218610120. Epub 2023 May 1"
COMMENT     Sequence Label: Ydf NanoLuc
FEATURES             Location/Qualifiers
     source          1..3826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(49..153)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     221..239
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(277..299)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     399..418
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     612..634
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     626..643
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     627..643
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1372..1401
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1424..1453
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1469..1486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1542..1559)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1712..1758
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1854..1870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1854..1870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1867..1889)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1878..1894
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1902..1908,1909..1926,1927..1932))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1947..1968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2085..2102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2255..2843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2335..2354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3014..3805,3806..3826,1..48))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3637..3656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
       61 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
      121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
      181 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      241 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      301 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      361 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      421 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      481 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      541 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      601 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtaagatct cggctgaatt
      661 ctaaagatct actgcatatt tgaaaattgc ccaaacgtac atgcccgaat gtacgttttt
      721 ttcatttcat tgtcaactac aatgagaaag aatgtgatca agcaatgtgt tgaaaggaga
      781 ttatcacgtc gactctcgag tgagattgtt gacggtaccg tattttggat ctaggaggaa
      841 ggatctatgg tgtttacgct ggaggatttc gtcggtgact ggcgtcagac agctgggtat
      901 aaccttgacc aggtacttga acaaggcggc gtttccagct tatttcaaaa tctgggggtg
      961 tctgtcacac caattcagcg cattgtcttg tctggggaga atggtcttaa aattgatatt
     1021 catgttatca tcccttacga aggtttgtcc ggtgaccaaa tgggacaaat cgagaaaatt
     1081 ttcaaagtgg tttatccagt ggatgatcat cactttaaag tcattttaca ctatggtacg
     1141 ctggtaatcg acggtgtgac accgaacatg attgattatt tcgggcgtcc gtatgaagga
     1201 atcgccgttt ttgatgggaa gaaaatcacc gtaacaggta ctctgtggaa cggaaacaaa
     1261 atcatcgacg aacgcttgat taaccccgat ggtagtttgt tgttccgtgt tacgattaac
     1321 ggagttacgg gttggcgcct ttgtgagcgt atcttggccg gaagcggaag cgagcagaaa
     1381 ctcatctctg aagaggatct gtaaggatct gaagcttggg cccgaacaaa aactcatctc
     1441 agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag tttaaacggt
     1501 ctccagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     1561 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     1621 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     1681 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1741 ctgggccttt cgttttatct gttgtttgtc ggtgaactct gaaaggagga actatatccg
     1801 gatcgagatc ctctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag
     1861 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1921 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1981 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2041 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2101 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2161 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaagg
     2221 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2281 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2341 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2401 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2461 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2521 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2581 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2641 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2701 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2761 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2821 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2881 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2941 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3001 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3061 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3121 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3181 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3241 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3301 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3361 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3421 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3481 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3541 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3601 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3661 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3721 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3781 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaa
//
