LOCUS       sequence-189593-        1944 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 1944)
  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 1944)
  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: YCe2730 HC_Kan_conn_BsaI-4_p25
FEATURES             Location/Qualifiers
     source          1..1944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      287..316
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      338..925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     826..845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(937..964)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(988..1803)
                     /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     1711..1730
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1804..1906)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1907..1938)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tatggaggtc aggtatgatt taaatggtca gtattgagcg atatctagag aattcgtcac
       61 tatcctaacg tccacttcag cgtctcgact gtgcttcttt acgtagtagt tccatcccac
      121 ttatcgtccg cgagacccta atttgtgcat tgtcggcctt cgtccttctt tgttgctctg
      181 gctacgacga gacgcaggtg gatgaaagcg tgtaaacgaa ttcaagcttg atatcattca
      241 ggacgagcct cagactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg
      301 ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg
      361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      421 aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac
      481 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
      661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
      721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
      781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc
      901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg
      961 agccaggtga ttacatttgg gccctcatta gaaaaactca tcgagcatca agtgaaactg
     1021 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     1081 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     1141 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     1201 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     1261 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcacc
     1321 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgccgtt
     1381 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     1441 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttccctgg
     1501 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     1561 aagaggcata aattccgtca gccagtttag cctgaccatc tcatctgtaa catcattggc
     1621 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     1681 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     1741 agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct
     1801 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     1861 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     1921 ggtcggaggc ttttgacttt ctgc
//
