LOCUS       sequence-331007-        2201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of PbrR in E. coli for cell-free lead sensing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2201)
  AUTHORS   Jung JK, Rasor BJ, Rybnicky GA, Silverman AD, Standeven J, Kuhn R, 
            Granito T, Ekas HM, Wang BM, Karim AS, Lucks JB, Jewett MC
  TITLE     At-Home, Cell-Free Synthetic Biology Education Modules for 
            Transcriptional Regulation and Environmental Water Quality 
            Monitoring.
  JOURNAL   ACS Synth Biol. 2023 Oct 20;12(10):2909-2921. doi: 
            10.1021/acssynbio.3c00223. Epub 2023 Sep 12.
  PUBMED    37699423
REFERENCE   2  (bases 1 to 2201)
  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: 
            "10.1021/acssynbio.3c00223"; journalName: "ACS Synth Biol"; date: 
            "2023-10-20- 20"; volume: "12"; issue: "10"; pages: "2909-2921"
COMMENT     Sequence Label: pJBL7044
FEATURES             Location/Qualifiers
     source          1..2201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             683..705
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1202..1220)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1216..1263
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1281..2096)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2004..2023
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2088..2107
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg atcccgcgaa attaatacga ctcactatag ggagaccaca
      661 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgaacat
      721 tcagatcggt gaactggcga aacgtactgc atgtccagtg gtgaccattc gcttttatga
      781 gcaagaaggt ttgctgccgc ccccaggacg ttcccgtggt aatttccgct tatacgggga
      841 agagcatgtt gagcgtttgc aatttatccg ccattgccgc tcacttgaca tgcccttatc
      901 cgatgtccgc acattgctgt cataccgcaa gcgtcctgat caagattgcg gggaggtgaa
      961 catgcttctg gatgagcata ttcgtcaagt agagagtcgt attggcgctc tgttagagtt
     1021 aaaacatcac ttggtcgagc tgcgcgaagc atgctcaggg gctcgccctg cacaaagctg
     1081 cggaatcctt cagggcctga gtgattgtgt ttgcgataca cgtggcacga cagcacaccc
     1141 ctcggattaa gtcgaccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1201 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1261 ttgctgaaag ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     1321 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     1381 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1441 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1501 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     1561 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     1621 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     1681 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1741 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     1801 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1861 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1921 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     1981 gtagcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     2041 atgttggaat ttaatcgcgg cttcgagcaa gacgtttccc gttgaatatg gctcataaca
     2101 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     2161 tcttgtgcaa tgtaacatca gagattttga gacacaacgt g
//
