LOCUS       sequence-192914-        2409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  part designed to occupy position 21 of EMMA. Functional category: 
            Selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2409)
  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 2409)
  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: YCe2040 HC_Kan_PuroR_p21
FEATURES             Location/Qualifiers
     source          1..2409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      29..60
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        61..163
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             164..979
                     /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(237..256)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1000..1027
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1039..1626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1119..1138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1648..1677)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(1765..2361)
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(1848..1868)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 taaatcatac ctgacctcca tagcagaaag tcaaaagcct ccgaccggag gcttttgact
       61 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
      121 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatgagcc atattcaacg
      181 ggaaacgtct tgctcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
      241 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
      301 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
      361 agatgagatg gtcaggctaa actggctgac ggaatttatg cctcttccga ccatcaagca
      421 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatcccag ggaaaacagc
      481 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
      541 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaacg gcgatcgcgt
      601 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttggtg cgagtgattt
      661 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
      721 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
      781 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
      841 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
      901 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcacttgat
      961 gctcgatgag tttttctgag ggcccaaatg taatcacctg gctcaccttc gggtgggcct
     1021 ttctgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1081 atgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1141 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1201 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1261 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1321 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1381 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1441 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1501 tctgctgaag ccagttacct cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1561 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1621 tctcaagaag atcctttgat tttctaccga agaaaggccc acccgtgaag gtgagccagt
     1681 gagttgattg cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt
     1741 gaattcgttt cgtctcgcaa cttattaggc accgggcttg cgggtcatgc accaggtcgc
     1801 gcggtccttc gggcactcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg
     1861 gaggttgcgg ggcgcggagg tttccaggaa ggcgggcacc ccggcgcgct cggccgcctc
     1921 cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgacacgcc
     1981 gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat
     2041 ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc
     2101 gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc
     2161 gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag
     2221 ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc
     2281 ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg
     2341 caccgtgggc ttgtactcgg ttccacgaga cgtgacgaat tctctagata tcgctcaata
     2401 ctgaccatt
//
