LOCUS       sequence-296938-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides Middle-to-End linker 2 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 2
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2..59
                     /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(83..100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(254..842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(334..353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1025..1119)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1140..1955)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1863..1882
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2117..2135
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2139..2182)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
       61 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      241 cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      961 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc
     1021 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     1081 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
     1141 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     1201 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     1261 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     1321 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     1381 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     1441 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     1501 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     1561 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     1621 tcttctaata cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca
     1681 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     1741 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     1801 aactctggcg catcgggctt cccatacaat ccatagattg tcgcacctga ttgcccgaca
     1861 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     1921 ctggagcaag acgtttcccg ttgaatatgg ctcattttag cttccttagc tcctgaaaat
     1981 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     2041 cctcttacgt gcccgatcaa ctcgagtgcc acctgacgtc taagaaacca ttattatcat
     2101 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
     2161 agctttcgct aaggatgatt tctggaattc cacctgcata tgtcacgcta gagaccgtat
     2221 gaggtggatg gtgtgctgcg tggccaaacc catatgcagg tgctgca
//
