LOCUS       sequence-314735-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides Middle-to-End linker 1 for level 2 cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2267)
  AUTHORS   Andreou , Andreas I., Nirkko, Jessica , Ochoa-Villarreal, Marisol , 
            Nakayama, Naomi
  TITLE     Mobius Assembly for Plant Systems highlights promoter-terminator 
            interaction in gene regulation
  JOURNAL   bioRxiv, March 31, 2021
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: "bioRxiv, 
            March 31, 2021"
COMMENT     Sequence Label: Auxiliary 1
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      18..75
                     /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(99..116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(270..858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(350..369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1041..1135)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1156..1971)
                     /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(1156..1971)
                     /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     1879..1898
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2133..2151
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2155..2198)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 atatgcaggt gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct
       61 ttaaaaccga aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc
      121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      181 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      241 aaccgtaaaa aggccgcgtt gctggcgttt ttccacaggc tccgcccccc tgacgagcat
      301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
      721 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
      781 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      841 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
      901 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
      961 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1021 tctgacagct cgaggcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     1081 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     1141 gagctcgata tcaaattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1201 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1261 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1321 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1381 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     1441 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     1501 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     1561 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     1621 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgcagtggt
     1681 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1741 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1801 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcggt agattgtcgc
     1861 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1921 ggaatttaat cgcggccttg agcaagacgt ttcccgttga atatggctca ttttagcttc
     1981 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     2041 atggtgaaag ttggaacctc ttacgtgccc gatcaactcg agtgccacct gacgtctaag
     2101 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca
     2161 gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattccacc tgcatatcag
     2221 acgctagaga ccgtatgagg tggatggtgt gctgcgtggc caaaccc
//
