LOCUS       sequence-192749-        1848 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 1848)
  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 1848)
  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..1848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      98..127
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      149..736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     637..656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(748..775)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(796..1611)
                     /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     1519..1538
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1612..1714)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1715..1746)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tcactaaagg ctaagtcccg agacgtacga attcaagctt gatatcattc aggacgagcc
       61 tcagactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
      121 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
      181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      241 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
      301 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      361 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      421 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      481 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      541 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      601 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      661 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
      721 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
      781 attacatttg ggccctcaga aaaactcatc gagcatcaag tgaaactgca atttattcat
      841 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
      901 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
      961 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1021 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     1081 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcaccaa ccaaaccgtt
     1141 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgccgttaa aaggacaatt
     1201 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     1261 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgcagtggt
     1321 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1381 ttccgtcagc cagtttagcc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1441 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     1501 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1561 ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca ttttagcttc
     1621 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     1681 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     1741 ttgactttct gctatggagg tcaggtatga tttaaatggt cagtattgag cgatatctag
     1801 agaattcgtc gtctcgcgga tagatcgatc tgaattaatt ctgacggt
//
