LOCUS       sequence-324767-        2171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of CueR in E. coli for cell-free copper sensing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2171)
  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 2171)
  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: pJBL7045
FEATURES             Location/Qualifiers
     source          1..2171
                     /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(1172..1190)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1186..1233
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1251..2066)
                     /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     1974..1993
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2058..2077
                     /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 atatgaatat
      721 ctctgatgtt gcaaagatta cgggccttac ctcgaaagca atccgcttct acgaagaaaa
      781 gggactggtc acacctccca tgcgtagcga aaacggctac cgcacataca cccagcaaca
      841 cctgaatgag ctgactttgc tgcgccaggc gcgtcaagtg ggtttcaatt tggaggaatc
      901 cggggagctg gtcaacctgt tcaatgaccc acaacgccat tcggccgacg tgaaacgccg
      961 tacattagaa aaagttgccg aaattgaacg ccatattgaa gagctgcagt ccatgcgcga
     1021 tcagttgtta gcactggcaa acgcttgtcc tggagatgat tcggctgatt gtccgattat
     1081 cgaaaactta tctggatgtt gtcaccatcg cgcagggtaa gtcgaccggc tgctaacaaa
     1141 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     1201 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag ccaattctga ttagaaaaac
     1261 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1321 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1381 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1441 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1501 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     1561 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     1621 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     1681 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1741 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     1801 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1861 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1921 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     1981 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cttcgagcaa
     2041 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     2101 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     2161 gacacaacgt g
//
