LOCUS       sequence-189560-        1852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit Assembly connector D-E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1852)
  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 1852)
  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: YCe2024 HC_Kan_conn_D-E
FEATURES             Location/Qualifiers
     source          1..1852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      129..160
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        161..263
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             264..1079
                     /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(337..356)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1103..1130
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1142..1729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1222..1241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1751..1780)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 acgtctcggg acttagcctt tagtgaaccg tcagaattaa ttcagatcga tctatccgcg
       61 agacgacgaa ttctctagat atcgctcaat actgaccatt taaatcatac ctgacctcca
      121 tagcagaaag tcaaaagcct ccgaccggag gcttttgact tgatcggcac gtaagaggtt
      181 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
      241 tttcaggagc taaggaagct aaaatgagcc atattcaacg ggaaacgtct tgctcgaggc
      301 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
      361 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
      421 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcaggctaa
      481 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
      541 atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta ttagaagaat
      601 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
      661 cgattcctgt ttgtaattgt ccttttaacg gcgatcgcgt atttcgtctc gctcaggcgc
      721 aatcacgaat gaataacggt ttggttggtg cgagtgattt tgatgacgag cgtaatggct
      781 ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag
      841 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
      901 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
      961 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1021 ttgataatcc tgatatgaat aaattgcagt ttcacttgat gctcgatgag tttttctaat
     1081 gagggcccaa atgtaatcac ctggctcacc ttcgggtggg cctttctgcg ttgctggcgt
     1141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgatgctca agtcagaggt
     1201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1561 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1621 cctcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1741 gattttctac cgaagaaagg cccacccgtg aaggtgagcc agtgagttga ttgcagtcca
     1801 gttacgctgg agtctgaggc tcgtcctgaa tgatatcaag cttgaattcg tt
//
