LOCUS       sequence-324768-        2225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of CadR in E. coli for cell-free cadmium sensing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2225)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 2225)
  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"
COMMENT     Sequence Label: pJBL7047
FEATURES             Location/Qualifiers
     source          1..2225
                     /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)"
     RBS             713..724
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1226..1244)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1240..1287
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1305..2120)
                     /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     2028..2047
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2112..2131
                     /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 ataaagagga
      721 gaaatactag atgaaaatcg gtgaacttgc taaagccacg gactgtgcgg ttgaaactat
      781 ccgttactac gagcgtgaac agttgctgcc agagcctgcc cgttcagacg gtaattatcg
      841 cttatacacg caggcacatg tggaacgtct gacgtttatt cgcaactgcc gtacattgga
      901 tatgactctt gatgaaatcc gttctctgtt gcgtttgcgt gattcgccgg atgatagctg
      961 cgggtccgtg aacgcgttaa ttgatgaaca tattgagcat gtgcaagctc gtatcgatgg
     1021 actggttgcg cttcaggagc aactggtaga acttcgtcgt cgctgcaatg cccaaggtgc
     1081 tgagtgtgcc atcttacagc aattggagac gaacggtgct gtttcggtcc ccgaaacaga
     1141 acactcccat gtgggtcgct ctcacggcca ctaggtcgac cggctgctaa caaagcccga
     1201 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     1261 tctaaacggg tcttgagggg ttttttgctg aaagccaatt ctgattagaa aaactcatcg
     1321 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     1381 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     1441 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1501 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1561 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     1621 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     1681 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1741 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1801 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1861 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1921 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1981 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     2041 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcttcga gcaagacgtt
     2101 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     2161 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     2221 acgtg
//
