LOCUS       sequence-227536-        2555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of aadA (streptomycin resistance gene) from the 
            T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2555)
  AUTHORS   Stark JC, Huang A, Hsu KJ, Dubner RS, Forbrook J, Marshalla S, 
            Rodriguez F, Washington M, Rybnicky GA, Nguyen PQ, Hasselbacher B, 
            Jabri R, Kamran R, Koralewski V, Wightkin W, Martinez T, Jewett MC
  TITLE     BioBits Health: Classroom Activities Exploring Engineering, Biology,
            and Human Health with Fluorescent Readouts.
  JOURNAL   ACS Synth Biol. 2019 May 7. doi: 10.1021/acssynbio.8b00381.
  PUBMED    30925042
REFERENCE   2  (bases 1 to 2555)
  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"; journalName: "ACS Synth 
            Biol. 2019 May 7. doi: 10.1021/acssynbio.8b00381."
COMMENT     Sequence Label: pJL1-aadA
FEATURES             Location/Qualifiers
     source          1..2555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      112..700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     601..620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     728..747
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        728..746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             778..800
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             808..1599
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(1651..1669)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1665..1712
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1730..2545)
                     /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     2453..2472
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(2537..2555,1)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
       61 gcaatgtaac atcagagatt ttgagacaca acgtgagatc aaaggatctt cttgagatcc
      121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      241 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcgatccc
      721 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt
      781 gtttaacttt aagaaggaga tatacatatg agggaagcgg tgatcgccga agtatcgact
      841 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
      901 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
      961 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
     1021 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
     1081 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
     1141 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
     1201 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
     1261 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
     1321 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
     1381 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
     1441 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
     1501 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
     1561 tacgtgaaag gcgagatcac caaggtagtc ggcaaataag tcgaccggct gctaacaaag
     1621 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1681 gggcctctaa acgggtcttg aggggttttt tgctgaaagc caattctgat tagaaaaact
     1741 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1801 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1861 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     1921 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     1981 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2041 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2101 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2161 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2221 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2281 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2341 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2401 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2461 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcgagcaag
     2521 acgtttcccg ttgaatatgg ctcataacac ccctt
//
