LOCUS       sequence-196914-        1908 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 1908)
  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 1908)
  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: YCe4520 HC_Kan_conn_BsaI-1_p1
FEATURES             Location/Qualifiers
     source          1..1908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      188..219
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        220..322
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             323..1138
                     /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(396..415)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1159..1186
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1198..1785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1278..1297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1807..1836)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 agtcaccgtc tcgtaggcgg aacacgcgaa gcgaagcaga ggtctcagga cagtacatgt
       61 aaccatagtg tcaagaacag tcgcgacgat ggagtaagat taagggtcaa tggcgagacg
      121 ctagacgaat tctctagata tcgctcaata ctgaccattt aaatcatacc tgacctccat
      181 agcagaaagt caaaagcctc cgaccggagg cttttgactt gatcggcacg taagaggttc
      241 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
      301 ttcaggagct aaggaagcta aaatgagcca tattcaacgg gaaacgtctt gctcgaggcc
      361 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
      421 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
      481 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcaggctaaa
      541 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
      601 tgcatggtta ctcaccactg cgatcccagg gaaaacagca ttccaggtat tagaagaata
      661 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
      721 gattcctgtt tgtaattgtc cttttaacgg cgatcgcgta tttcgtctcg ctcaggcgca
      781 atcacgaatg aataacggtt tggttggtgc gagtgatttt gatgacgagc gtaatggctg
      841 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
      901 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
      961 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1021 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1081 tgataatcct gatatgaata aattgcagtt tcacttgatg ctcgatgagt ttttctgagg
     1141 gcccaaatgt aatcacctgg ctcaccttcg ggtgggcctt tctgcgttgc tggcgttttt
     1201 ccataggctc cgcccccctg acgagcatca caaaaatcga tgctcaagtc agaggtggcg
     1261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctc
     1681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatt
     1801 ttctaccgaa gaaaggccca cccgtgaagg tgagccagtg agttgattgc agtccagtta
     1861 cgctggagtc tgaggctcgt cctgaatgat atcaagcttg aattcgtt
//
