LOCUS       sequence-252952-       10571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level1 ZmUBIp::FnCas12a (rice codon optimized):NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10571)
  AUTHORS   Hahn F, Korolev A, Sanjurjo Loures L, Nekrasov V
  TITLE     A modular cloning toolkit for genome editing in plants.
  JOURNAL   BMC Plant Biol. 2020 Apr 23;20(1):179. doi: 
            10.1186/s12870-020-02388-2.
  PUBMED    32326901
REFERENCE   2  (bases 1 to 10571)
  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: "BMC Plant 
            Biol."; date: "2020-04-23"; volume: "20(1):179. doi"; pages: " 
            10.1186/s12870-020-02388-2"
COMMENT     Sequence Label: pFH60
FEATURES             Location/Qualifiers
     source          1..10571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5938..5958
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5974..6226
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6274..6298
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(6454..7602)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(7938..8526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8018..8037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8697..9488,9489..9557))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9320..9339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9558..9662)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9749..10380
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    10472..10496
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caagcaagaa ttcaagcttg gaggtgcagc gtgacccggt cgtgcccctc
       61 tctagagata atgagcattg catgtctaag ttataaaaaa ttaccacata ttttttttgt
      121 cacacttgtt tgaagtgcag tttatctatc tttatacata tatttaaact ttactctacg
      181 aataatataa tctatagtac tacaataata tcagtgtttt agagaatcat ataaatgaac
      241 agttagacat ggtctaaagg acaattgagt attttgacaa caggactcta cagttttatc
      301 tttttagtgt gcatgtgttc tccttttttt ttgcaaatag cttcacctat ataatacttc
      361 atccatttta ttagtacatc catttagggt ttagggttaa tggtttttat agactaattt
      421 ttttagtaca tctattttat tctattttag cctctaaatt aagaaaacta aaactctatt
      481 ttagtttttt tatttaataa tttagatata aaatagaata aaataaagtg actaaaaatt
      541 aaacaaatac cctttaagaa attaaaaaaa ctaaggaaac atttttcttg tttcgagtag
      601 ataatgccag cctgttaaac gccgtcgacg agtctaacgg acaccaacca gcgaaccagc
      661 agcgtcgcgt cgggccaagc gaagcagacg gcacggcatc tctgtcgctg cctctggacc
      721 cctctcgaga gttccgctcc accgttggac ttgctccgct gtcggcatcc agaaattgcg
      781 tggcggagcg gcagacgtga gccggcacgg caggcggcct cctcctcctc tcacggcacg
      841 gcagctacgg gggattcctt tcccaccgct ccttcgcttt cccttcctcg cccgccgtaa
      901 taaatagaca ccccctccac accctctttc cccaacctcg tgttgttcgg agcgcacaca
      961 cacacaacca gatctccccc aaatccaccc gtcggcacct ccgcttcaag gtacgccgct
     1021 cgtcctcccc ccccccccct ctctaccttc tctagatcgg cgttccggtc catggttagg
     1081 gcccggtagt tctacttctg ttcatgtttg tgttagatcc gtgtttgtgt tagatccgtg
     1141 ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac acgttctgat tgctaacttg
     1201 ccagtgtttc tctttgggga atcctgggat ggctctagcc gttccgcaga cgggatcgat
     1261 ttcatgattt tttttgtttc gttgcatagg gtttggtttg cccttttcct ttatttcaat
     1321 atatgccgtg cacttgtttg tcgggtcatc ttttcatgct tttttttgtc ttggttgtga
     1381 tgatgtggtc tggttgggcg gtcgttctag atcggagtag aattctgttt caaactacct
     1441 ggtggattta ttaattttgg atctgtatgt gtgtgccata catattcata gttacgaatt
     1501 gaagatgatg gatggaaata tcgatctagg ataggtatac atgttgatgc gggttttact
     1561 gatgcatata cagagatgct ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg
     1621 gtcgttcatt cgttctagat cggagtagaa tactgtttca aactacctgg tgtatttatt
     1681 aattttggaa ctgtatgtgt gtgtcataca tcttcatagt tacgagttta agatggatgg
     1741 aaatatcgat ctaggatagg tatacatgtt gatgtgggtt ttactgatgc atatacatga
     1801 tggcatatgc agcatctatt catatgctct aaccttgagt acctatctat tataataaac
     1861 aagtatgttt tataattatt ttgatcttga tatacttgga tgatggcata tgcagcagct
     1921 atatgtggat ttttttagcc ctgccttcat acgctattta tttgcttggt actgtttctt
     1981 ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag gaatgtccat ctaccaagag
     2041 ttcgtgaaca agtactccct gtccaagacc ctccgcttcg agcttatccc acagggcaag
     2101 acgctcgaga acatcaaggc caggggcctc atcctcgacg acgagaagag agccaaggac
     2161 tacaagaagg ccaagcagat catcgacaag taccaccagt tctttatcga ggagatcctc
     2221 tcctccgtct gcatctccga ggacctgctc cagaactact ccgacgtgta cttcaagctc
     2281 aagaagtccg acgacgacaa cctccagaag gacttcaagt ccgccaagga caccatcaag
     2341 aagcagatct ccgagtacat taaggactcc gagaagttca agaacctctt caaccagaac
     2401 ctcatcgacg ccaagaaggg gcaagagtcc gacctcatcc tgtggctcaa gcagtccaag
     2461 gacaacggca tcgagctgtt caaggccaac tccgacatca ccgacatcga cgaggccctc
     2521 gagatcatta agtccttcaa gggctggacc acctacttca aggggttcca cgagaaccgc
     2581 aagaacgtgt actcctccaa cgacatcccg acctccatca tctacaggat cgtggacgat
     2641 aacctgccga agttccttga gaacaaggcc aagtacgagt ccctcaagga caaggcccca
     2701 gaggccatca actacgagca gatcaagaag gacctcgccg aggagctgac cttcgacatc
     2761 gattacaaga cctccgaggt gaaccagcgc gtgttctcac tcgacgaggt gttcgagatc
     2821 gccaacttca acaactacct caaccagtcc ggcatcacca agttcaacac catcatcggc
     2881 ggcaagttcg tcaacggcga gaacacaaag cgcaagggca tcaacgagta catcaacctc
     2941 tactcccagc agattaacga caagaccctg aagaagtaca agatgtccgt cctcttcaag
     3001 caaatcctct ccgacaccga gtccaagtcc ttcgtgatcg acaagctcga ggacgacagc
     3061 gacgtggtga ccaccatgca gtccttctac gagcaaatcg ccgccttcaa gaccgtcgag
     3121 gagaagtcca tcaaggagac tctcagcctc ctgttcgacg acctcaaggc ccagaagctc
     3181 gacctctcca agatctattt caagaacgat aagtccctca ccgacctcag ccagcaggtt
     3241 ttcgacgact actccgtgat cggcacagcc gtgctcgagt atatcacgca gcagatcgcc
     3301 ccgaagaacc tcgacaaccc gagcaagaag gagcaagagc tgatcgccaa aaagaccgag
     3361 aaggcgaagt acctcagcct cgagacaatc aagctcgccc tcgaggagtt caacaagcac
     3421 cgcgacattg acaagcagtg ccgctttgag gagattctcg cgaacttcgc cgcgatcccg
     3481 atgatcttcg atgagatcgc ccagaacaag gataacctcg cccagatctc catcaagtac
     3541 cagaaccagg ggaagaagga tcttctccag gcctccgccg aggacgatgt gaaggctatt
     3601 aaggacctcc tcgaccagac caacaacctc ctccacaagc ttaagatctt ccacatctcc
     3661 cagtccgagg ataaggccaa catcctggac aaggacgagc acttctacct cgtgttcgag
     3721 gagtgctact tcgagctggc gaacatcgtg ccgctctaca acaagatccg caactacatc
     3781 acccagaagc cgtactcgga cgagaagttt aagctcaact tcgagaactc caccctcgcc
     3841 aacggctggg acaagaacaa ggagccagac aacaccgcca tcctcttcat caaggatgac
     3901 aagtactacc tcggcgtgat gaacaagaag aacaacaaga ttttcgacga taaggcgatc
     3961 aaggagaaca agggcgaggg ctacaagaag atcgtctaca agctcctccc aggcgccaac
     4021 aagatgctcc cgaaggtgtt cttcagcgcc aagagcatca agttctacaa cccgtccgag
     4081 gacatcctcc gcatccgcaa ccattccacc cacaccaaga atggctcccc gcagaagggc
     4141 tacgagaagt tcgagttcaa catcgaggac tgccgcaagt tcatcgactt ctacaagcag
     4201 agcatctcca agcacccaga gtggaaggat ttcggcttcc gcttcagcga cacccagcgc
     4261 tacaacagca tcgatgagtt ctaccgcgag gtcgagaatc agggctataa gctgacgttc
     4321 gagaatatct ccgagagcta catcgactcc gtggtcaacc agggcaagct ctacctgttc
     4381 cagatctata acaaggattt ctccgcctac agcaagggca ggccgaacct ccataccctc
     4441 tactggaagg ccctcttcga cgagaggaac ctccaggacg tggtgtacaa gctcaatggc
     4501 gaggcggagc tgttctaccg gaagcagtct atcccgaaga agatcaccca cccggccaag
     4561 gaggccattg ccaacaagaa caaggacaac ccgaagaagg agtccgtttt cgagtacgac
     4621 ctgatcaagg ataagcgctt taccgaggac aagttcttct tccactgccc gatcaccatc
     4681 aacttcaaga gcagcggcgc gaacaagttc aatgacgaga tcaacctcct gctgaaggag
     4741 aaggctaacg acgtccacat cctctccatc gacaggggcg agagacacct cgcctactac
     4801 acactcgtgg acggcaaggg caacatcatc aagcaggaca ccttcaatat catcggcaac
     4861 gaccgcatga aaaccaacta ccacgataag ctggccgcca tcgagaagga cagggattcc
     4921 gctaggaagg actggaagaa gattaacaat attaaggaga tgaaggaggg ctacctctcc
     4981 caggtggtgc acgagattgc caagctcgtc atcgagtaca acgccatcgt ggttttcgag
     5041 gaccttaact tcggcttcaa gcgcggcagg ttcaaggttg agaagcaggt ctaccaaaag
     5101 ctcgagaaga tgctcattga gaagctcaat tacctggtgt ttaaggataa cgagttcgac
     5161 aagacaggcg gcgtgctcag agcctaccaa ctcacagctc cgtttgagac attcaagaag
     5221 atgggcaagc agaccggcat tatctactac gtgccagccg gcttcaccag caagatctgc
     5281 ccagtgaccg gcttcgtcaa ccagctctac ccgaagtacg agagcgtgtc caagtcgcaa
     5341 gagttcttta gcaagttcga taagatctgc tacaacctcg ataagggcta ctttgagttc
     5401 agcttcgact ataagaattt cggcgacaag gccgcgaagg gcaagtggac aattgcttcc
     5461 ttcggctccc gcctgatcaa cttccgcaac agcgacaaga accacaactg ggacacccgc
     5521 gaggtgtacc ccacaaagga gttggagaag ctcctgaagg actactccat tgagtacggc
     5581 cacggcgagt gcatcaaggc tgccatttgc ggcgagtccg ataagaagtt cttcgccaag
     5641 ctcacctccg tgctcaacac gatcctccag atgcgcaact ccaagaccgg caccgagctg
     5701 gactacctca tttccccagt ggccgacgtg aacggcaact tcttcgattc taggcaggcc
     5761 ccaaagaaca tgccgcagga tgccgatgcc aacggcgcct atcatattgg cctcaagggc
     5821 ctcatgctcc tgggccgcat caagaacaat caagagggca agaagctcaa cctcgtcatc
     5881 aagaacgagg agtacttcga gttcgtgcag aaccggaaca acgactccag ggccgaccca
     5941 aagaagaaga ggaaggtctg agcttgtcaa gcagatcgtt caaacatttg gcaataaagt
     6001 ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat
     6061 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
     6121 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca
     6181 aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcgacg ctactattgt
     6241 cttctgcacg aagtggttta aactatcagt gtttgacagg atatattggc gggtaaacct
     6301 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
     6361 cgttcgtcca tttgtatgtg catgccaacc acagggttcc ccagatcagg cgctggctgc
     6421 tgaaccccca gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc
     6481 attgacccag gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac
     6541 ctgctcgcgc cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc
     6601 cagcttgagc gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt
     6661 cggcgacagc ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag
     6721 cacgacgatt tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag
     6781 gacgcggaag cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa
     6841 gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc
     6901 attgatcgac cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc
     6961 gataggggtg cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc
     7021 ccgcagctcg acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt
     7081 gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc
     7141 cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag
     7201 ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct
     7261 gacctgtttt gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg
     7321 cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc tgttcaagac gacgcgaacg
     7381 ctccacggcg gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc
     7441 gatcttggcc gtagcttgct ggaccatcga gccgacggac tggaaggttt cgcggggcgc
     7501 acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat
     7561 cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat
     7621 tgccccgact cacgccgggg caatgtgccc ttattcctga tttgacccgc ctggtgcctt
     7681 ggtgtccaga taatccacct tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc
     7741 cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa
     7801 atacttgccg tgggcctcgg cctgagagcc aaaacacttg atgcggaaga agtcggtgcg
     7861 ctcctgcttg tcgccggcat cgttgcgcca catctaggat ctgccaggaa ccgtaaaaag
     7921 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7981 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     8041 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     8101 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     8161 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     8221 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     8281 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     8341 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     8401 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8461 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8521 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8581 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8641 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8701 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8761 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8821 gctgcaatga taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag
     8881 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8941 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     9001 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     9061 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     9121 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     9181 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9241 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     9301 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9361 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9421 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9481 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9541 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcaaggttat
     9601 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9661 cgcacgaatt ggccagcgct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca
     9721 gcccctgggg ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca
     9781 ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt
     9841 ataaatattg gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg
     9901 aaacccttgc aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc
     9961 gcccctcatc tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca
    10021 gtagtcgcgc ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc
    10081 atctgtggga aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc
    10141 cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg
    10201 gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg
    10261 tatccacaac gccggcggcc gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg
    10321 tttgcagggc catagacggc cgccagccca gcggcgaggg caaccagccc ggtgagcgtc
    10381 gcaaaggaga tcctgatctg actgatgggc tgcctgtatc gagtggtgat tttgtgccga
    10441 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
    10501 tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggatgca
    10561 gtgggcccca c
//
