LOCUS       sequence-189591-        1943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit Assembly connector for second step assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1943)
  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 1943)
  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: YCe2725 HC_Kan_conn_BsaI-2_p24
FEATURES             Location/Qualifiers
     source          1..1943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      220..251
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        252..354
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             355..1170
                     /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(428..447)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1194..1221
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1233..1820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1313..1332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1842..1871)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 actatcctaa cgtccacttc agcgtctcgc acgatcacca cctaaactcc aacgcaatat
       61 ctaaccacaa caatctcact ctatcccgag acccctccca acccataagg ctcgtccaat
      121 ctcaaactgc gagacgcagg tggatgaaag cgtgtaacga attctctaga tatcgctcaa
      181 tactgaccat ttaaatcata cctgacctcc atagcagaaa gtcaaaagcc tccgaccgga
      241 ggcttttgac ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca
      301 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatgagc
      361 catattcaac gggaaacgtc ttgctcgagg ccgcgattaa attccaacat ggatgctgat
      421 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
      481 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
      541 aatgatgtta cagatgagat ggtcaggcta aactggctga cggaatttat gcctcttccg
      601 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatccca
      661 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
      721 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
      781 ggcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttggt
      841 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
      901 cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
      961 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     1021 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     1081 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     1141 tttcacttga tgctcgatga gtttttctaa tgagggccca aatgtaatca cctggctcac
     1201 cttcgggtgg gcctttctgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1261 catcacaaaa atcgatgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1681 tttggtatct gcgctctgct gaagccagtt acctcggaaa aagagttggt agctcttgat
     1741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1801 gcagaaaaaa aggatctcaa gaagatcctt tgattttcta ccgaagaaag gcccacccgt
     1861 gaaggtgagc cagtgagttg attgcagtcc agttacgctg gagtctgagg ctcgtcctga
     1921 atgatatcaa gcttgaattc gtt
//
