LOCUS       sequence-314912-        2853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Confers resistance to kanamycin and G418.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2853)
  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 2853)
  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: lvl0 NptII
FEATURES             Location/Qualifiers
     source          1..2853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..97)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(251..839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(331..350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1022..1116)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1137..1796)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1711..1730
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1958..1976
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1980..2023)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             2040..2834
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2094..2113)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2704..2723
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      join(2852..2853,1..56)
                     /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."
ORIGIN
        1 cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac
       61 ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      241 tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc tcgaggcttg
     1021 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     1081 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac
     1141 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     1201 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     1261 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     1321 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     1381 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     1441 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     1501 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     1561 tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     1621 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     1681 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     1741 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     1801 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     1861 tcattatggt gaaagttgga acctcttacg tgcccgatca actcgagtgc cacctgacgt
     1921 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa
     1981 tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt cggtctcaaa
     2041 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2101 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2161 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactcc
     2221 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2281 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2341 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2401 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     2461 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     2521 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgg atgcccgacg
     2581 gcgaggatct cgtcgtgacc cacggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     2641 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     2701 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     2761 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     2821 acgagttctt ctgagcttcg agaccctgca gtc
//
