LOCUS       sequence-192757-        1869 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 1869)
  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 1869)
  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..1869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      214..245
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        246..348
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             349..1164
                     /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(422..441)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1185..1212
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1224..1811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1304..1323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1833..1862)
                     /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 gccgtttatg acgaattctc tagatatcgc tcaatactga
      181 ccatttaaat catacctgac ctccatagca gaaagtcaaa agcctccgac cggaggcttt
      241 tgacttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
      301 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat gagccatatt
      361 caacgggaaa cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat
      421 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
      481 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
      541 gttacagatg agatggtcag gctaaactgg ctgacggaat ttatgcctct tccgaccatc
      601 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat cccagggaaa
      661 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
      721 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacggcgat
      781 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tggtgcgagt
      841 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag
      901 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
      961 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     1021 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     1081 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcac
     1141 ttgatgctcg atgagttttt ctgagggccc aaatgtaatc acctggctca ccttcgggtg
     1201 ggcctttctg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1261 aatcgatgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1621 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1681 tgcgctctgc tgaagccagt tacctcggaa aaagagttgg tagctcttga tccggcaaac
     1741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1801 aaggatctca agaagatcct ttgattttct accgaagaaa ggcccacccg tgaaggtgag
     1861 ccagtgagt
//
