LOCUS       sequence-314737-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides Middle-to-End linker 3 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 3
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5..62
                     /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(86..103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(257..845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(337..356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1028..1122)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1143..1958)
                     /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(1143..1958)
                     /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     1866..1885
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2120..2138
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2142..2185)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gcagtccggc aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
       61 gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      121 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      181 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      241 ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac aaaaatcgac
      301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
      901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagctcga
     1021 ggcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     1081 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca
     1141 aattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1201 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1261 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1321 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1381 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1441 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1501 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1561 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1621 tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag taaccatgca
     1681 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1741 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1801 aacaactctg gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc tgattgcccg
     1861 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1921 ggccttgagc aagacgtttc ccgttgaata tggctcattt tagcttcctt agctcctgaa
     1981 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     2041 gaacctctta cgtgcccgat caactcgagt gccacctgac gtctaagaaa ccattattat
     2101 catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc
     2161 cttagctttc gctaaggatg atttctggaa ttccacctgc atatcttgcg ctagagaccg
     2221 tatgaggtgg atggtgtgct gcgtggccaa acccatatgc aggtgct
//
