LOCUS       sequence-314738-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides End-to-End linker 4A 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 A
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      25..82
                     /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(106..123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(277..865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(357..376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1048..1142)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1163..1978)
                     /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(1163..1978)
                     /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     1886..1905
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2140..2158
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2162..2205)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 caaacccata tgcaggtgct gcagtccggc aaaaaagggc aaggtgtcac caccctgccc
       61 tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc
      121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc gcccccctga
      301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1021 aacttggtct gacagctcga ggcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     1081 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     1141 tccaagcgag ctcgatatca aattagaaaa actcatcgag catcaaatga aactgcaatt
     1201 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     1261 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     1321 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1381 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     1441 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     1501 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     1561 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     1621 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg
     1681 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1741 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     1801 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcggtaga
     1861 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1921 ccatgttgga atttaatcgc ggccttgagc aagacgtttc ccgttgaata tggctcattt
     1981 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     2041 tttcattatg gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac
     2101 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag
     2161 aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttccacctgc
     2221 atatcgctca gaagagaccg tatgaggtgg atggtgtgct gcgtggc
//
