LOCUS       sequence-314739-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides End-to-End linker 4B 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 B
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      20..77
                     /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(101..118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(272..860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(352..371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1043..1137)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1158..1973)
                     /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(1158..1973)
                     /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     1881..1900
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2135..2153
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2157..2200)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 ccatatgcag gtgctgcagt ccggcaaaaa agggcaaggt gtcaccaccc tgcccttttt
       61 ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc
      121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag gctccgcccc cctgacgagc
      301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      781 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
      901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
      961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1021 ggtctgacag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     1081 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     1141 gcgagctcga tatcaaatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     1201 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     1261 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     1321 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     1381 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     1441 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     1501 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     1561 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1621 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttccctgg gatcgcagtg
     1681 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1741 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1801 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg gtagattgtc
     1861 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1921 ttggaattta atcgcggcct tgagcaagac gtttcccgtt gaatatggct cattttagct
     1981 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     2041 ttatggtgaa agttggaacc tcttacgtgc ccgatcaact cgagtgccac ctgacgtcta
     2101 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt
     2161 cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcca cctgcatatc
     2221 gctgtcaaga gaccgtatga ggtggatggt gtgctgcgtg gccaaac
//
