LOCUS       sequence-237143-        2901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coding sequence nptII from E. coli domesticated into pUPD2 with 
            AATG/GCTT barcodes. According to FungalBraid/GoldenBraid modular DNA
            assembly for A. tumefaciens-mediated genetic transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2901)
  AUTHORS   Hernanz-Koers M, Gandia M, Garrigues S, Manzanares P, Yenush L, 
            Orzaez D, Marcos JF
  TITLE     FungalBraid: A GoldenBraid-based modular cloning platform for the 
            assembly and exchange of DNA elements tailored to fungal synthetic 
            biology.
  JOURNAL   Fungal Genet Biol. 2018 Jul;116:51-61. doi: 
            10.1016/j.fgb.2018.04.010. Epub 2018 Apr 20.
  PUBMED    29680684
REFERENCE   2  (bases 1 to 2901)
  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: "Fungal 
            Genet Biol."; date: "2018-07"; volume: "116:51-61. doi"; pages: " 
            10.1016/j.fgb.2018.04.010. Epub 2018 Apr 20"
COMMENT     Sequence Label: FB005
FEATURES             Location/Qualifiers
     source          1..2901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     297..316
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      469..526
                     /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(550..567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(721..1309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(801..820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1492..1586)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1607..2266)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2181..2200
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2428..2446
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2450..2493)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2496..2517
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             join(2534..2901,1..427)
                     /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(2588..2607)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacacc gcatcgagcg
       61 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
      121 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cggatgcccg acggcgagga
      181 tctcgtcgtg acccacggcg acgcctgctt gccgaatatc atggtggaaa atggccgctt
      241 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
      301 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
      361 ttacggtatc gccgctcccg attcgcagcg catagccttc tatcgccttc ttgacgagtt
      421 cttctgagct ttgagaccac tcatcgcgac tagtagcggc cgctgcagtc cggcaaaaaa
      481 gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat
      541 gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      601 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      661 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      721 tttccacagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      781 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      841 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      901 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      961 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1021 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1081 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1141 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1201 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1261 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1321 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1381 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1441 atcaatctaa agtatatatg agtaaacttg gtctgacagc tcgaggcttg gattctcacc
     1501 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     1561 cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct
     1621 gccactcgtc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     1681 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     1741 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     1801 aactggtgaa actcacccag ggattggctg atacgaaaaa catattctca ataaaccctt
     1861 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     1921 actgccggaa atcgtcgtgg tattcactcc agagtgatga aaacgtttca gtttgctcat
     1981 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     2041 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     2101 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     2161 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     2221 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     2281 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     2341 gaaagttgga acctcttacg tgcccgatca actcgagtgc cacctgacgt ctaagaaacc
     2401 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa
     2461 aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct tctagagcga
     2521 tgagtggtct cgaatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2581 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2641 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2701 cctgaatgaa ctccaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2761 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2821 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2881 ggctgatgca atgcggcggc t
//
