LOCUS       sequence-229836-        2579 bp    DNA     circular SYN 15-APR-2025
DEFINITION  In vitro expression of neoR (kanamycin resistance gene) from the T7 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2579)
  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 2579)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-neoR
FEATURES             Location/Qualifiers
     source          1..2579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      67..655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     556..575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     683..702
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        683..701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             733..755
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             763..1578
                     /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     complement(1630..1648)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1644..1691
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1709..2524)
                     /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     2516..2535
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg agatcaaagg
       61 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      121 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      181 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      241 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      301 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      361 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      421 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      481 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      541 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      601 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      661 cagcaacgcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
      721 tctagaaata attttgttta actttaagaa ggagatatac atatgagcca tattcaacgg
      781 gaaacgtctt gctcgaagcc gcgattaaat tccaacatgg atgctgattt atatgggtat
      841 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
      901 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
      961 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1021 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca
     1081 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1141 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1201 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1261 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     1321 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     1381 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     1441 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     1501 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     1561 ctcgatgagt ttttctaagt cgaccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     1621 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     1681 ggggtttttt gctgaaagcc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     1741 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     1801 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     1861 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     1921 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     1981 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     2041 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     2101 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     2161 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     2221 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     2281 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     2341 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     2401 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     2461 cagcatccat gttggaattt aatcgcggct tcgagcaaga cgtttcccgt tgaatatggc
     2521 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgat
//
