LOCUS       sequence-314747-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides End-to-End linker 4-Delta 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 Delta
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      14..71
                     /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(95..112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(266..854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(346..365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1037..1131)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1152..1967)
                     /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(1152..1967)
                     /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     1875..1894
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2129..2147
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2151..2194)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gcaggtgctg cagtccggca aaaaagggca aggtgtcacc accctgccct ttttctttaa
       61 aaccgaaaag attacttcgc gttatgcagg cttcctcgct cactgactcg ctgcgctcgg
      121 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      181 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      241 gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca
      301 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      361 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      421 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      481 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      541 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      601 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      661 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      721 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      781 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      841 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      901 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      961 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1021 acagctcgag gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     1081 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     1141 tcgatatcaa attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1201 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1261 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1321 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1381 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     1441 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1501 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     1561 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     1621 gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc agtggtgagt
     1681 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     1741 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     1801 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcggtagat tgtcgcacct
     1861 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     1921 tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcatttt agcttcctta
     1981 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2041 tgaaagttgg aacctcttac gtgcccgatc aactcgagtg ccacctgacg tctaagaaac
     2101 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata
     2161 aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tccacctgca tatcgctaga
     2221 gaccttctgt atgaggtgga tggtgtgctg cgtggccaaa cccatat
//
