LOCUS       sequence-456402-        2717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 promoter driving expression of metR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2717)
  AUTHORS   Piorino F, Johnson S, Styczynski MP
  TITLE     A Cell-Free Biosensor for Assessment of Hyperhomocysteinemia.
  JOURNAL   ACS Synth Biol. 2023 Aug 18;12(8):2487-2492. doi: 
            10.1021/acssynbio.3c00103. Epub 2023 Jul 17.
  PUBMED    37459448
REFERENCE   2  (bases 1 to 2717)
  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.3c00103"; journalName: "ACS Synth Biol"; date: 
            "2023-08-18- 18"; volume: "12"; issue: "8"; pages: "2487-2492"
COMMENT     Sequence Label: pMetR
FEATURES             Location/Qualifiers
     source          1..2717
                     /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(1718..1736)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1732..1779
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1797..2612)
                     /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     2520..2539
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2604..2623
                     /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 atatgatcga
      721 agtaaaacac ctgaaaacgc tacaagcgtt gcggaactgc ggctcgctcg cagccgctgc
      781 ggcgacgttg catcagacgc aatccgccct gtctcaccag tttagcgatc tggaacaacg
      841 ccttggcttc cggctatttg tgcgtaagag ccagccgcta cgctttacac cgcagggaga
      901 aatcctgttg caactggcaa accaggtact gccgcaaatt agccaggccc tgcaagcctg
      961 caatgaaccg cagcagacgc gtctgcgcat tgccattgag tgccatagct gtattcagtg
     1021 gctgacaccc gcgttagaaa atttccataa gaactggccg caggtagaga tggattttaa
     1081 atcgggcgtg acatttgacc cgcagcccgc cttgcaacag ggagagctgg atctggtaat
     1141 gacgtccgat attctgccgc gcagtggcct gcattattcg ccgatgttcg actatgaagt
     1201 gcgtctggtg ttagcacctg accatccact ggcggcgaaa acgcgaatta caccggaaga
     1261 tctcgccagc gagacgctat taatttatcc ggtgcagcgt agtcgactgg atgtctggcg
     1321 gcattttctt cagccggcag gcgtcagccc gtcactgaaa agcgtcgata acaccttatt
     1381 gttgattcag atggttgccg cgcggatggg tattgccgcg ctaccgcatt gggtagtaga
     1441 gagttttgag cgccagggtc tggtggtgac caaaaccctg ggcgaaggct tgtggagccg
     1501 actgtacgcc gccgtgcgcg atggcgagca gcgtcagcca gttacggaag cgtttattcg
     1561 ctcagcgcgc aatcacgcct gcgatcatct gccgtttgtg aagagcgcgg agcgacccac
     1621 ttacgatgca cccacagtga ggccaggatc accagcgcgc ctgtaagtcg accggctgct
     1681 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1741 ccacttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaagccaa ttctgattag
     1801 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     1861 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     1921 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     1981 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     2041 tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca
     2101 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     2161 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     2221 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     2281 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     2341 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     2401 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     2461 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     2521 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcttc
     2581 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     2641 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     2701 ttttgagaca caacgtg
//
