LOCUS       sequence-189563-        1871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit Assembly connector B-E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1871)
  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 1871)
  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: YCe2982 HC_Kan_conn_B-E
FEATURES             Location/Qualifiers
     source          1..1871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      213..244
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        245..347
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             348..1163
                     /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(421..440)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1187..1214
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1226..1813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1306..1325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1835..1864)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tgattgcagt ccagttacgc tggagtctga ggctcgtcct gaatgatatc aagcttgaat
       61 tcgtttttaa cgcagccgtc tccatggact aactacggtg aaagagactt agaaggagat
      121 tgagagatcc gtgagacgag gccgttatga cgaattctct agatatcgct caatactgac
      181 catttaaatc atacctgacc tccatagcag aaagtcaaaa gcctccgacc ggaggctttt
      241 gacttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
      301 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc
      361 aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg
      421 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
      481 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
      541 ttacagatga gatggtcagg ctaaactggc tgacggaatt tatgcctctt ccgaccatca
      601 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa
      661 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
      721 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacggcgatc
      781 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt ggtgcgagtg
      841 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
      901 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
      961 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     1021 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     1081 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcact
     1141 tgatgctcga tgagtttttc taatgagggc ccaaatgtaa tcacctggct caccttcggg
     1201 tgggcctttc tgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1261 aaaatcgatg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1681 tctgcgctct gctgaagcca gttacctcgg aaaaagagtt ggtagctctt gatccggcaa
     1741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1801 aaaaggatct caagaagatc ctttgatttt ctaccgaaga aaggcccacc cgtgaaggtg
     1861 agccagtgag t
//
