LOCUS       sequence-189617-        2423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  part designed to occupy position 15 of EMMA. Functional category: 
            Selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2423)
  AUTHORS   Martella A, Matjusaitis M, Auxillos J, Pollard SM, Cai Y
  TITLE     EMMA: An Extensible Mammalian Modular Assembly Toolkit for the Rapid
            Design and Production of Diverse Expression Vectors.
  JOURNAL   ACS Synth Biol. 2017 Apr 24. doi: 10.1021/acssynbio.7b00016.
  PUBMED    28418644
REFERENCE   2  (bases 1 to 2423)
  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. 2017 Apr 24. doi: 10.1021/acssynbio.7b00016."
COMMENT     Sequence Label: YCe2046 HC_Kan_PuroR_p15
FEATURES             Location/Qualifiers
     source          1..2423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             89..688
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(89..108)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     585..605
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     terminator      768..799
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        800..902
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             903..1718
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(976..995)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1742..1769
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1781..2368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1861..1880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2390..2419)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 agttgattgc agtccagtta cgctggagtc tgaggctcgt cctgaatgat atcaagcttg
       61 aattcgttac gtctcggcaa ccgccaccat gaccgagtac aagcccacgg tgcgcctcgc
      121 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
      181 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
      241 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
      301 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
      361 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
      421 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtgtc
      481 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
      541 cgagcgcgcc ggggtgcccg ccttcctgga aacctccgcg ccccgcaacc tccccttcta
      601 cgagcggctc ggcttcaccg tcaccgccga cgtcgagtgc ccgaaggacc gcgcgacctg
      661 gtgcatgacc cgcaagcccg gtgcctaata accctcgaga cgaaacgaat tctctagata
      721 tcgctcaata ctgaccattt aaatcatacc tgacctccat agcagaaagt caaaagcctc
      781 cgaccggagg cttttgactt gatcggcacg taagaggttc caactttcac cataatgaaa
      841 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
      901 aaatgagcca tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg
      961 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     1021 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1081 gcgttgccaa tgatgttaca gatgagatgg tcaggctaaa ctggctgacg gaatttatgc
     1141 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     1201 cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     1261 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     1321 cttttaacgg cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     1381 tggttggtgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     1441 aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     1501 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     1561 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     1621 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     1681 aattgcagtt tcacttgatg ctcgatgagt ttttctaatg agggcccaaa tgtaatcacc
     1741 tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg ctccgccccc
     1801 ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg acaggactat
     1861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2221 gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa gagttggtag
     2281 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2341 gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc gaagaaaggc
     2401 ccacccgtga aggtgagcca gtg
//
