LOCUS       sequence-271525-       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             4893..4940
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4956..5208
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5256..5280
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(5436..6584)
                     /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(6920..7508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7000..7019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7679..8470,8471..8539))
                     /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     8302..8321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8540..8644)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8731..9362
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    9454..9478
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ccccccccct ctctaccttc tctagatcgg cgttccggtc catggttagg gcccggtagt
       61 tctacttctg ttcatgtttg tgttagatcc gtgtttgtgt tagatccgtg ctgctagcgt
      121 tcgtacacgg atgcgacctg tacgtcagac acgttctgat tgctaacttg ccagtgtttc
      181 tctttgggga atcctgggat ggctctagcc gttccgcaga cgggatcgat ttcatgattt
      241 tttttgtttc gttgcatagg gtttggtttg cccttttcct ttatttcaat atatgccgtg
      301 cacttgtttg tcgggtcatc ttttcatgct tttttttgtc ttggttgtga tgatgtggtc
      361 tggttgggcg gtcgttctag atcggagtag aattctgttt caaactacct ggtggattta
      421 ttaattttgg atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg
      481 gatggaaata tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata
      541 cagagatgct ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt
      601 cgttctagat cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa
      661 ctgtatgtgt gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat
      721 ctaggatagg tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc
      781 agcatctatt catatgctct aaccttgagt acctatctat tataataaac aagtatgttt
      841 tataattatt ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat
      901 ttttttagcc ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc
      961 tcaccctgtt gtttggtgtt acttctgcag gaatgagcat ctaccaggag ttcgtcaaca
     1021 agtattcact gagtaagaca ctgcggttcg agctgatccc acagggcaag acactggaga
     1081 acatcaaggc ccgaggcctg attctggacg atgagaagcg ggcaaaagac tataagaaag
     1141 ccaagcagat cattgataaa taccaccagt tctttatcga ggaaattctg agctccgtgt
     1201 gcatcagtga ggatctgctg cagaattact cagacgtgta cttcaagctg aagaagagcg
     1261 acgatgacaa cctgcagaag gacttcaagt ccgccaagga caccatcaag aaacagatta
     1321 gcgagtacat caaggactcc gaaaagttta aaaatctgtt caaccagaat ctgatcgatg
     1381 ctaagaaagg ccaggagtcc gacctgatcc tgtggctgaa acagtctaag gacaatggga
     1441 ttgaactgtt caaggctaac tccgatatca ctgatattga cgaggcactg gaaatcatca
     1501 agagcttcaa gggatggacc acatacttta aaggcttcca cgagaaccgc aagaacgtgt
     1561 actccagcaa cgacattcct acctccatca tctaccgaat cgtcgatgac aatctgccaa
     1621 agttcctgga gaacaaggcc aaatatgaat ctctgaagga caaagctccc gaggcaatta
     1681 attacgaaca gatcaagaaa gatctggctg aggaactgac attcgatatc gactataaga
     1741 ctagcgaggt gaaccagagg gtcttttccc tggacgaggt gtttgaaatc gccaatttca
     1801 acaattacct gaaccagtcc ggcattacta aattcaatac catcattggc gggaagtttg
     1861 tgaacgggga gaataccaag cgcaagggaa ttaacgaata catcaatctg tatagccagc
     1921 agatcaacga caaaactctg aagaaataca agatgtctgt gctgttcaaa cagatcctga
     1981 gtgataccga gtccaagtct tttgtcattg ataaactgga agatgactca gacgtggtca
     2041 ctaccatgca gagcttttat gagcagatcg ccgctttcaa gacagtggag gaaaaatcta
     2101 ttaaggaaac tctgagtctg ctgttcgatg acctgaaagc ccagaagctg gacctgagta
     2161 agatctactt caaaaacgat aagagtctga cagacctgtc acagcaggtg tttgatgact
     2221 attccgtgat tgggaccgcc gtcctggagt acattacaca gcagatcgct ccaaagaacc
     2281 tggataatcc ctctaagaaa gagcaggaac tgatcgctaa gaaaaccgag aaggcaaaat
     2341 atctgagtct ggaaacaatt aagctggcac tggaggagtt caacaagcac agggatattg
     2401 acaaacagtg ccgctttgag gaaatcctgg ccaacttcgc agccatcccc atgatttttg
     2461 atgagatcgc ccagaacaaa gacaatctgg ctcagatcag tattaagtac cagaaccagg
     2521 gcaagaaaga cctgctgcag gcttcagcag aagatgacgt gaaagccatc aaggatctgc
     2581 tggaccagac caacaatctg ctgcacaagc tgaaaatctt ccatattagt cagtcagagg
     2641 ataaggctaa tatcctggat aaagacgaac acttctacct ggtgttcgag gaatgttact
     2701 tcgagctggc aaacattgtc cccctgtata acaagattag gaactacatc acacagaagc
     2761 cttactctga cgagaagttt aaactgaact tcgaaaatag taccctggcc aacgggtggg
     2821 ataagaacaa ggagcctgac aacacagcta tcctgttcat caaggatgac aagtactatc
     2881 tgggagtgat gaataagaaa aacaataaga tcttcgatga caaagccatt aaggagaaca
     2941 aaggggaagg atacaagaaa atcgtgtata agctgctgcc cggcgcaaat aagatgctgc
     3001 ctaaggtgtt cttcagcgcc aagagtatca aattctacaa cccatccgag gacatcctgc
     3061 ggattagaaa tcactcaaca catactaaga acgggagccc ccagaaggga tatgagaaat
     3121 ttgagttcaa catcgaggat tgcaggaagt ttattgactt ctacaagcag agcatctcca
     3181 aacaccctga atggaaggat tttggcttcc ggttttccga cacacagaga tataactcta
     3241 tcgacgagtt ctaccgcgag gtggaaaatc aggggtataa gctgactttt gagaacattt
     3301 ctgaaagtta catcgacagc gtggtcaatc agggaaagct gtacctgttc cagatctata
     3361 acaaagattt ttcagcatac agcaagggca gaccaaacct gcatacactg tactggaagg
     3421 ccctgttcga tgagaggaat ctgcaggacg tggtctataa actgaacgga gaggccgaac
     3481 tgttttaccg gaagcagtct attcctaaga aaatcactca cccagctaag gaggccatcg
     3541 ctaacaagaa caaggacaat cctaagaaag agagcgtgtt cgaatacgat ctgattaagg
     3601 acaagcggtt caccgaagat aagttctttt tccattgtcc aatcaccatt aacttcaagt
     3661 caagcggcgc taacaagttc aacgacgaga tcaatctgct gctgaaggaa aaagcaaacg
     3721 atgtgcacat cctgagcatt gaccgaggag agcggcatct ggcctactat accctggtgg
     3781 atggcaaagg gaatatcatt aagcaggata cattcaacat cattggcaat gaccggatga
     3841 aaaccaacta ccacgataaa ctggctgcaa tcgagaagga tagagactca gctaggaagg
     3901 actggaagaa aatcaacaac attaaggaga tgaaggaagg ctatctgagc caggtggtcc
     3961 atgagattgc aaagctggtc atcgaataca atgccattgt ggtgttcgag gatctgaact
     4021 tcggctttaa gagggggcgc tttaaggtgg aaaaacaggt ctatcagaag ctggagaaaa
     4081 tgctgatcga aaagctgaat tacctggtgt ttaaagataa cgagttcgac aagaccggag
     4141 gcgtcctgag agcctaccag ctgacagctc cctttgaaac tttcaagaaa atgggaaaac
     4201 agacaggcat catctactat gtgccagccg gattcacttc caagatctgc cccgtgaccg
     4261 gctttgtcaa ccagctgtac cctaaatatg agtcagtgag caagtcccag gaatttttca
     4321 gcaagttcga taagatctgt tataatctgg acaaggggta cttcgagttt tccttcgatt
     4381 acaagaactt cggcgacaag gccgctaagg ggaaatggac cattgcctcc ttcggatctc
     4441 gcctgatcaa ctttcgaaat tccgataaaa accacaattg ggacactagg gaggtgtacc
     4501 caaccaagga gctggaaaag ctgctgaaag actactctat cgagtatgga catggcgaat
     4561 gcatcaaggc agccatctgt ggcgagagtg ataagaaatt tttcgccaag ctgacctcag
     4621 tgctgaatac aatcctgcag atgcggaact caaagaccgg gacagaactg gactatctga
     4681 ttagccccgt ggctgatgtc aacggaaact tcttcgacag cagacaggca cccaaaaata
     4741 tgcctcagga tgcagacgcc aacggggcct accacatcgg gctgaaggga ctgatgctgc
     4801 tgggccggat caagaacaat caggagggga agaagctgaa cctggtcatt aagaacgagg
     4861 aatacttcga gtttgtccag aatagaaata acaaaaggcc ggcggccacg aaaaaggccg
     4921 gccaggcaaa aaagaaaaag tgagcttgtc aagcagatcg ttcaaacatt tggcaataaa
     4981 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     5041 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     5101 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     5161 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcga cgctactatt
     5221 gtcttctgca cgaagtggtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
     5281 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
     5341 tccgttcgtc catttgtatg tgcatgccaa ccacagggtt ccccagatca ggcgctggct
     5401 gctgaacccc cagccggaac tgaccccaca aggccctagc gtttgcaatg caccaggtca
     5461 tcattgaccc aggcgtgttc caccaggccg ctgcctcgca actcttcgca ggcttcgccg
     5521 acctgctcgc gccacttctt cacgcgggtg gaatccgatc cgcacatgag gcggaaggtt
     5581 tccagcttga gcgggtacgg ctcccggtgc gagctgaaat agtcgaacat ccgtcgggcc
     5641 gtcggcgaca gcttgcggta cttctcccat atgaatttcg tgtagtggtc gccagcaaac
     5701 agcacgacga tttcctcgtc gatcaggacc tggcaacggg acgttttctt gccacggtcc
     5761 aggacgcgga agcggtgcag cagcgacacc gattccaggt gcccaacgcg gtcggacgtg
     5821 aagcccatcg ccgtcgcctg taggcgcgac aggcattcct cggccttcgt gtaataccgg
     5881 ccattgatcg accagcccag gtcctggcaa agctcgtaga acgtgaaggt gatcggctcg
     5941 ccgatagggg tgcgcttcgc gtactccaac acctgctgcc acaccagttc gtcatcgtcg
     6001 gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct tgttgacgtg gaaaatgacc
     6061 ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg tggtgaacag ggcagagcgg
     6121 gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg gcgcaatatc gaacaaggaa
     6181 agctgcattt ccttgatctg ctgcttcgtg tgtttcagca acgcggcctg cttggcctcg
     6241 ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct tcttggtcgt catagttcct
     6301 cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct cctgttcaag acgacgcgaa
     6361 cgctccacgg cggccgatgg cgcgggcagg gcagggggag ccagttgcac gctgtcgcgc
     6421 tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg actggaaggt ttcgcggggc
     6481 gcacgcatga cggtgcggct tgcgatggtt tcggcatcct cggcggaaaa ccccgcgtcg
     6541 atcagttctt gcctgtatgc cttccggtca aacgtccgat tcattcaccc tccttgcggg
     6601 attgccccga ctcacgccgg ggcaatgtgc ccttattcct gatttgaccc gcctggtgcc
     6661 ttggtgtcca gataatccac cttatcggca atgaagtcgg tcccgtagac cgtctggccg
     6721 tccttctcgt acttggtatt ccgaatcttg ccctgcacga ataccagcga ccccttgccc
     6781 aaatacttgc cgtgggcctc ggcctgagag ccaaaacact tgatgcggaa gaagtcggtg
     6841 cgctcctgct tgtcgccggc atcgttgcgc cacatctagg atctgccagg aaccgtaaaa
     6901 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6961 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7021 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7081 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7141 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7201 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7261 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7321 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7381 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7441 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7501 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7561 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7621 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7801 gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca gcaataaacc
     7861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7981 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8281 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcaaggtt
     8581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8641 cgcgcacgaa ttggccagcg ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg
     8701 cagcccctgg ggggatggga ggcccgcgtt agcgggccgg gagggttcga gaaggggggg
     8761 cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta aaaacaaggt
     8821 ttataaatat tggtttaaaa gcaggttaaa agacaggtta gcggtggccg aaaaacgggc
     8881 ggaaaccctt gcaaatgctg gattttctgc ctgtggacag cccctcaaat gtcaataggt
     8941 gcgcccctca tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt
     9001 cagtagtcgc gcccctcaag tgtcaatacc gcagggcact tatccccagg cttgtccaca
     9061 tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag gctggccagc
     9121 tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt
     9181 cggcccctca agtgtcaacg tccgcccctc atctgtcagt gagggccaag ttttccgcga
     9241 ggtatccaca acgccggcgg ccgcggtgtc tcgcacacgg cttcgacggc gtttctggcg
     9301 cgtttgcagg gccatagacg gccgccagcc cagcggcgag ggcaaccagc ccggtgagcg
     9361 tcgcaaagga gatcctgatc tgactgatgg gctgcctgta tcgagtggtg attttgtgcc
     9421 gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa
     9481 attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact ggggtggatg
     9541 cagtgggccc cactctgtga agacaagcaa gaattcaagc ttggaggtgc agcgtgaccc
     9601 ggtcgtgccc ctctctagag ataatgagca ttgcatgtct aagttataaa aaattaccac
     9661 atattttttt tgtcacactt gtttgaagtg cagtttatct atctttatac atatatttaa
     9721 actttactct acgaataata taatctatag tactacaata atatcagtgt tttagagaat
     9781 catataaatg aacagttaga catggtctaa aggacaattg agtattttga caacaggact
     9841 ctacagtttt atctttttag tgtgcatgtg ttctcctttt tttttgcaaa tagcttcacc
     9901 tatataatac ttcatccatt ttattagtac atccatttag ggtttagggt taatggtttt
     9961 tatagactaa tttttttagt acatctattt tattctattt tagcctctaa attaagaaaa
    10021 ctaaaactct attttagttt ttttatttaa taatttagat ataaaataga ataaaataaa
    10081 gtgactaaaa attaaacaaa taccctttaa gaaattaaaa aaactaagga aacatttttc
    10141 ttgtttcgag tagataatgc cagcctgtta aacgccgtcg acgagtctaa cggacaccaa
    10201 ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag acggcacggc atctctgtcg
    10261 ctgcctctgg acccctctcg agagttccgc tccaccgttg gacttgctcc gctgtcggca
    10321 tccagaaatt gcgtggcgga gcggcagacg tgagccggca cggcaggcgg cctcctcctc
    10381 ctctcacggc acggcagcta cgggggattc ctttcccacc gctccttcgc tttcccttcc
    10441 tcgcccgccg taataaatag acaccccctc cacaccctct ttccccaacc tcgtgttgtt
    10501 cggagcgcac acacacacaa ccagatctcc cccaaatcca cccgtcggca cctccgcttc
    10561 aaggtacgcc gctcgtcctc ccc
//
