LOCUS       sequence-201182-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the Middle-to-End linker 2 which assists Level 2 cloning in
            Mobius Assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2267)
  AUTHORS   Andreou AI, Nakayama N
  TITLE     Mobius Assembly: A versatile Golden-Gate framework towards universal
            DNA assembly.
  JOURNAL   PLoS One. 2018 Jan 2;13(1):e0189892. doi: 
            10.1371/journal.pone.0189892. eCollection 2018.
  PUBMED    29293531
REFERENCE   2  (bases 1 to 2267)
  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: "PLoS One.";
            date: "2018-01-2"; volume: "13(1):e0189892. doi"; pages: " 
            10.1371/journal.pone.0189892. eCollection 2018"
COMMENT     Sequence Label: pMA_Auxiliary 2
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..58
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(82..99)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(253..841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(333..352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1024..1118)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1139..1954)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1139..1954)
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1862..1881
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2116..2134
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2138..2181)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
       61 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      241 gttgctggcg tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
      721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
      781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
      841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgaggct
     1021 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     1081 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     1141 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     1201 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     1261 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     1321 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     1381 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     1441 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     1501 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     1561 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     1621 cttctaatac ctggaatgct gttttccctg ggatcgcagt ggtgagtaac catgcatcat
     1681 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     1741 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     1801 actctggcgc atcgggcttc ccatacaatc ggtagattgt cgcacctgat tgcccgacat
     1861 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     1921 ttgagcaaga cgtttcccgt tgaatatggc tcattttagc ttccttagct cctgaaaatc
     1981 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     2041 ctcttacgtg cccgatcaac tcgagtgcca cctgacgtct aagaaaccat tattatcatg
     2101 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
     2161 gctttcgcta aggatgattt ctggaattcc acctgcatat gtcacgctag agaccgtatg
     2221 aggtggatgg tgtgctgcgt ggccaaaccc atatgcaggt gctgcag
//
