LOCUS       sequence-263424-       10583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level1 ZmUBIp::FnCas12a (human codon optimized):NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10583)
  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 10583)
  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: pFH132
FEATURES             Location/Qualifiers
     source          1..10583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5923..5970
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      5986..6238
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6286..6310
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(6466..7614)
                     /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(7950..8538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8030..8049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8709..9500,9501..9569))
                     /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     9332..9351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9570..9674)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9761..10392
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    10484..10508
                     /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 gaatgagcat ctaccaggag
     2041 ttcgtcaaca agtattcact gagtaagaca ctgcggttcg agctgatccc acagggcaag
     2101 acactggaga acatcaaggc ccgaggcctg attctggacg atgagaagcg ggcaaaagac
     2161 tataagaaag ccaagcagat cattgataaa taccaccagt tctttatcga ggaaattctg
     2221 agctccgtgt gcatcagtga ggatctgctg cagaattact cagacgtgta cttcaagctg
     2281 aagaagagcg acgatgacaa cctgcagaag gacttcaagt ccgccaagga caccatcaag
     2341 aaacagatta gcgagtacat caaggactcc gaaaagttta aaaatctgtt caaccagaat
     2401 ctgatcgatg ctaagaaagg ccaggagtcc gacctgatcc tgtggctgaa acagtctaag
     2461 gacaatggga ttgaactgtt caaggctaac tccgatatca ctgatattga cgaggcactg
     2521 gaaatcatca agagcttcaa gggatggacc acatacttta aaggcttcca cgagaaccgc
     2581 aagaacgtgt actccagcaa cgacattcct acctccatca tctaccgaat cgtcgatgac
     2641 aatctgccaa agttcctgga gaacaaggcc aaatatgaat ctctgaagga caaagctccc
     2701 gaggcaatta attacgaaca gatcaagaaa gatctggctg aggaactgac attcgatatc
     2761 gactataaga ctagcgaggt gaaccagagg gtcttttccc tggacgaggt gtttgaaatc
     2821 gccaatttca acaattacct gaaccagtcc ggcattacta aattcaatac catcattggc
     2881 gggaagtttg tgaacgggga gaataccaag cgcaagggaa ttaacgaata catcaatctg
     2941 tatagccagc agatcaacga caaaactctg aagaaataca agatgtctgt gctgttcaaa
     3001 cagatcctga gtgataccga gtccaagtct tttgtcattg ataaactgga agatgactca
     3061 gacgtggtca ctaccatgca gagcttttat gagcagatcg ccgctttcaa gacagtggag
     3121 gaaaaatcta ttaaggaaac tctgagtctg ctgttcgatg acctgaaagc ccagaagctg
     3181 gacctgagta agatctactt caaaaacgat aagagtctga cagacctgtc acagcaggtg
     3241 tttgatgact attccgtgat tgggaccgcc gtcctggagt acattacaca gcagatcgct
     3301 ccaaagaacc tggataatcc ctctaagaaa gagcaggaac tgatcgctaa gaaaaccgag
     3361 aaggcaaaat atctgagtct ggaaacaatt aagctggcac tggaggagtt caacaagcac
     3421 agggatattg acaaacagtg ccgctttgag gaaatcctgg ccaacttcgc agccatcccc
     3481 atgatttttg atgagatcgc ccagaacaaa gacaatctgg ctcagatcag tattaagtac
     3541 cagaaccagg gcaagaaaga cctgctgcag gcttcagcag aagatgacgt gaaagccatc
     3601 aaggatctgc tggaccagac caacaatctg ctgcacaagc tgaaaatctt ccatattagt
     3661 cagtcagagg ataaggctaa tatcctggat aaagacgaac acttctacct ggtgttcgag
     3721 gaatgttact tcgagctggc aaacattgtc cccctgtata acaagattag gaactacatc
     3781 acacagaagc cttactctga cgagaagttt aaactgaact tcgaaaatag taccctggcc
     3841 aacgggtggg ataagaacaa ggagcctgac aacacagcta tcctgttcat caaggatgac
     3901 aagtactatc tgggagtgat gaataagaaa aacaataaga tcttcgatga caaagccatt
     3961 aaggagaaca aaggggaagg atacaagaaa atcgtgtata agctgctgcc cggcgcaaat
     4021 aagatgctgc ctaaggtgtt cttcagcgcc aagagtatca aattctacaa cccatccgag
     4081 gacatcctgc ggattagaaa tcactcaaca catactaaga acgggagccc ccagaaggga
     4141 tatgagaaat ttgagttcaa catcgaggat tgcaggaagt ttattgactt ctacaagcag
     4201 agcatctcca aacaccctga atggaaggat tttggcttcc ggttttccga cacacagaga
     4261 tataactcta tcgacgagtt ctaccgcgag gtggaaaatc aggggtataa gctgactttt
     4321 gagaacattt ctgaaagtta catcgacagc gtggtcaatc agggaaagct gtacctgttc
     4381 cagatctata acaaagattt ttcagcatac agcaagggca gaccaaacct gcatacactg
     4441 tactggaagg ccctgttcga tgagaggaat ctgcaggacg tggtctataa actgaacgga
     4501 gaggccgaac tgttttaccg gaagcagtct attcctaaga aaatcactca cccagctaag
     4561 gaggccatcg ctaacaagaa caaggacaat cctaagaaag agagcgtgtt cgaatacgat
     4621 ctgattaagg acaagcggtt caccgaagat aagttctttt tccattgtcc aatcaccatt
     4681 aacttcaagt caagcggcgc taacaagttc aacgacgaga tcaatctgct gctgaaggaa
     4741 aaagcaaacg atgtgcacat cctgagcatt gaccgaggag agcggcatct ggcctactat
     4801 accctggtgg atggcaaagg gaatatcatt aagcaggata cattcaacat cattggcaat
     4861 gaccggatga aaaccaacta ccacgataaa ctggctgcaa tcgagaagga tagagactca
     4921 gctaggaagg actggaagaa aatcaacaac attaaggaga tgaaggaagg ctatctgagc
     4981 caggtggtcc atgagattgc aaagctggtc atcgaataca atgccattgt ggtgttcgag
     5041 gatctgaact tcggctttaa gagggggcgc tttaaggtgg aaaaacaggt ctatcagaag
     5101 ctggagaaaa tgctgatcga aaagctgaat tacctggtgt ttaaagataa cgagttcgac
     5161 aagaccggag gcgtcctgag agcctaccag ctgacagctc cctttgaaac tttcaagaaa
     5221 atgggaaaac agacaggcat catctactat gtgccagccg gattcacttc caagatctgc
     5281 cccgtgaccg gctttgtcaa ccagctgtac cctaaatatg agtcagtgag caagtcccag
     5341 gaatttttca gcaagttcga taagatctgt tataatctgg acaaggggta cttcgagttt
     5401 tccttcgatt acaagaactt cggcgacaag gccgctaagg ggaaatggac cattgcctcc
     5461 ttcggatctc gcctgatcaa ctttcgaaat tccgataaaa accacaattg ggacactagg
     5521 gaggtgtacc caaccaagga gctggaaaag ctgctgaaag actactctat cgagtatgga
     5581 catggcgaat gcatcaaggc agccatctgt ggcgagagtg ataagaaatt tttcgccaag
     5641 ctgacctcag tgctgaatac aatcctgcag atgcggaact caaagaccgg gacagaactg
     5701 gactatctga ttagccccgt ggctgatgtc aacggaaact tcttcgacag cagacaggca
     5761 cccaaaaata tgcctcagga tgcagacgcc aacggggcct accacatcgg gctgaaggga
     5821 ctgatgctgc tgggccggat caagaacaat caggagggga agaagctgaa cctggtcatt
     5881 aagaacgagg aatacttcga gtttgtccag aatagaaata acaaaaggcc ggcggccacg
     5941 aaaaaggccg gccaggcaaa aaagaaaaag tgagcttgtc aagcagatcg ttcaaacatt
     6001 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa
     6061 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     6121 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     6181 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcga
     6241 cgctactatt gtcttctgca cgaagtggtt taaactatca gtgtttgaca ggatatattg
     6301 gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg
     6361 aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt ccccagatca
     6421 ggcgctggct gctgaacccc cagccggaac tgaccccaca aggccctagc gtttgcaatg
     6481 caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca actcttcgca
     6541 ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc cgcacatgag
     6601 gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat agtcgaacat
     6661 ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg tgtagtggtc
     6721 gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg acgttttctt
     6781 gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt gcccaacgcg
     6841 gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct cggccttcgt
     6901 gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga acgtgaaggt
     6961 gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc acaccagttc
     7021 gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct tgttgacgtg
     7081 gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg tggtgaacag
     7141 ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg gcgcaatatc
     7201 gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca acgcggcctg
     7261 cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct tcttggtcgt
     7321 catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct cctgttcaag
     7381 acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag ccagttgcac
     7441 gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg actggaaggt
     7501 ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct cggcggaaaa
     7561 ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat tcattcaccc
     7621 tccttgcggg attgccccga ctcacgccgg ggcaatgtgc ccttattcct gatttgaccc
     7681 gcctggtgcc ttggtgtcca gataatccac cttatcggca atgaagtcgg tcccgtagac
     7741 cgtctggccg tccttctcgt acttggtatt ccgaatcttg ccctgcacga ataccagcga
     7801 ccccttgccc aaatacttgc cgtgggcctc ggcctgagag ccaaaacact tgatgcggaa
     7861 gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc cacatctagg atctgccagg
     7921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     8161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     8401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     8461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     8521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8821 tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca
     8881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     9421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     9481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     9541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     9601 tatcaaggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9661 ataggggttc cgcgcacgaa ttggccagcg ctgccatttt tggggtgagg ccgttcgcgg
     9721 ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg gagggttcga
     9781 gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta
     9841 aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta gcggtggccg
     9901 aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag cccctcaaat
     9961 gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc
    10021 cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact tatccccagg
    10081 cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag
    10141 gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt caacgccgcg
    10201 ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt gagggccaag
    10261 ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc tcgcacacgg cttcgacggc
    10321 gtttctggcg cgtttgcagg gccatagacg gccgccagcc cagcggcgag ggcaaccagc
    10381 ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg gctgcctgta tcgagtggtg
    10441 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
    10501 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
    10561 ggggtggatg cagtgggccc cac
//
