LOCUS       sequence-189565-        1899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EMMA toolkit Assembly connector H-K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1899)
  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 1899)
  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: YCe1921 HC_Kan_conn_H-K
FEATURES             Location/Qualifiers
     source          1..1899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      243..272
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      294..881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     782..801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(893..920)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(944..1759)
                     /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     1667..1686
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1760..1862)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1863..1894)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctatggaggt caggtatgat ttaaatggtc agtattgagc gatatctaga gaattcgtct
       61 gcatccacga tctagtcacc gtctcaaggc taataacagc ttccggactc tagaacatcc
      121 ctacaggtga tatcctcggg tacgagacgc tactcaccct cgagcataac aacgaattca
      181 agcttgatat cattcaggac gagcctcaga ctccagcgta actggactgc aatcaactca
      241 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
      301 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      361 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
      421 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      481 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      541 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      601 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      661 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      721 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      781 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
      841 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
      901 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcattagaaa aactcatcga
      961 gcatcaagtg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1021 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1081 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1141 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1201 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1261 caaaatcact cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     1321 atacgcgatc gccgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     1381 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     1441 atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     1501 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg accatctcat
     1561 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     1621 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     1681 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
     1741 cccgttgaat atggctcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     1801 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     1861 caagtcaaaa gcctccggtc ggaggctttt gactttctg
//
