LOCUS       sequence-192766-        1849 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 1849)
  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 1849)
  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..1849
                     /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      1101..1128
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1140..1727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1220..1239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1749..1778)
                     /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 gtttttctga
     1081 gggcccaaat gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt gctggcgttt
     1141 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gatgctcaag tcagaggtgg
     1201 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1261 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1321 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1381 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1441 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1501 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1561 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1741 ttttctaccg aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gcagtccagt
     1801 tacgctggag tctgaggctc gtcctgaatg atatcaagct tgaattcgt
//
