LOCUS       sequence-324792-        2687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive cell-free expression of XylE enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2687)
  TITLE     Cell-free sensor distribution for primary synthetic biology 
            education
REFERENCE   2  (bases 1 to 2687)
  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: pJBL7076
FEATURES             Location/Qualifiers
     source          1..2687
                     /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(1688..1706)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1702..1749
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1767..2582)
                     /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     2490..2509
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2574..2593
                     /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 atatgaacaa
      721 aggtgtaatg cgaccgggcc atgtgcagct gcgtgtactg gacatgagca aggccctgga
      781 acactacgtc gagttgctgg gcctgatcga gatggaccgt gacgaccagg gccgtgtcta
      841 tctgaaggct tggaccgaag tggataagtt ttccctggtg ctacgcgagg ctgacgagcc
      901 gggcatggat tttatgggtt tcaaggttgt ggatgaggat gctctccggc aactggagcg
      961 ggatctgatg gcatatggct gtgccgttga gcagctaccc gcaggtgaac tgaacagttg
     1021 tggccggcgc gtgcgcttcc aggccccctc cgggcatcac ttcgagttgt atgcagacaa
     1081 ggaatatact ggaaagtggg gtttgaatga cgtcaatccc gaggcatggc cgcgcgatct
     1141 gaaaggtatg gcggctgtgc gtttcgacca cgccctcatg tatggcgacg aattgccggc
     1201 gacctatgac ctgttcacca aggtgctcgg tttctatctg gccgaacagg tgctggacga
     1261 aaatggcacg cgcgtcgccc agtttctcag tctgtcgacc aaggcccacg acgtggcctt
     1321 cattcaccat ccggaaaaag gccgcctcca tcatgtgtcc ttccacctcg aaacctggga
     1381 agacttgctt cgcgccgccg acctgatctc catgaccgac acatctatcg atatcggccc
     1441 aacccgccac ggcctcactc acggcaagac catctacttc ttcgacccgt ccggtaaccg
     1501 caacgaagtg ttctgcgggg gagattacaa ctacccggac cacaaaccgg tgacctggac
     1561 caccgaccag ctgggcaagg cgatctttta ccacgaccgc attctcaacg aacgattcat
     1621 gaccgtgctg acctaagtcg accggctgct aacaaagccc gaaaggaagc tgagttggct
     1681 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1741 ggttttttgc tgaaagccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc
     1801 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     1861 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     1921 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     1981 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt
     2041 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     2101 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     2161 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     2221 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     2281 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     2341 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     2401 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     2461 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     2521 gcatccatgt tggaatttaa tcgcggcttc gagcaagacg tttcccgttg aatatggctc
     2581 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     2641 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtg
//
