LOCUS       sequence-189572-        1853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit Assembly connector J-K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1853)
  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 1853)
  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: YCe2022 HC_Kan_conn_J-K
FEATURES             Location/Qualifiers
     source          1..1853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(22..41)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     terminator      130..161
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        162..264
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             265..1080
                     /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(338..357)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1104..1131
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1143..1730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1223..1242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1752..1781)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 acgtctcgtg ctgaggctaa ctgaaacacg gaaggagaca atacaggaag gaacggtacg
       61 agacggacga attctctaga tatcgctcaa tactgaccat ttaaatcata cctgacctcc
      121 atagcagaaa gtcaaaagcc tccgaccgga ggcttttgac ttgatcggca cgtaagaggt
      181 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
      241 ttttcaggag ctaaggaagc taaaatgagc catattcaac gggaaacgtc ttgctcgagg
      301 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
      361 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
      421 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcaggcta
      481 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
      541 gatgcatggt tactcaccac tgcgatccca gggaaaacag cattccaggt attagaagaa
      601 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
      661 tcgattcctg tttgtaattg tccttttaac ggcgatcgcg tatttcgtct cgctcaggcg
      721 caatcacgaa tgaataacgg tttggttggt gcgagtgatt ttgatgacga gcgtaatggc
      781 tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca
      841 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
      901 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
      961 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     1021 attgataatc ctgatatgaa taaattgcag tttcacttga tgctcgatga gtttttctaa
     1081 tgagggccca aatgtaatca cctggctcac cttcgggtgg gcctttctgc gttgctggcg
     1141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgatgctc aagtcagagg
     1201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1621 acctcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1741 tgattttcta ccgaagaaag gcccacccgt gaaggtgagc cagtgagttg attgcagtcc
     1801 agttacgctg gagtctgagg ctcgtcctga atgatatcaa gcttgaattc gtt
//
