LOCUS       sequence-252931-       10792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level1 ZmUBIp::nScCas9 (wheat codon optimized):NOSt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10792)
  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 10792)
  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: pFH109
FEATURES             Location/Qualifiers
     source          1..10792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6159..6179
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6195..6447
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6495..6519
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(6675..7823)
                     /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(8159..8747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8239..8258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8918..9709,9710..9778))
                     /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     9541..9560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9779..9883)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9970..10601
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    10693..10717
                     /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 cccccccctc tctaccttct ctagatcggc gttccggtcc atggttaggg
     1081 cccggtagtt ctacttctgt tcatgtttgt gttagatccg tgtttgtgtt agatccgtgc
     1141 tgctagcgtt cgtacacgga tgcgacctgt acgtcagaca cgttctgatt gctaacttgc
     1201 cagtgtttct ctttggggaa tcctgggatg gctctagccg ttccgcagac gggatcgatt
     1261 tcatgatttt ttttgtttcg ttgcataggg tttggtttgc ccttttcctt tatttcaata
     1321 tatgccgtgc acttgtttgt cgggtcatct tttcatgctt ttttttgtct tggttgtgat
     1381 gatgtggtct ggttgggcgg tcgttctaga tcggagtaga attctgtttc aaactacctg
     1441 gtggatttat taattttgga tctgtatgtg tgtgccatac atattcatag ttacgaattg
     1501 aagatgatgg atggaaatat cgatctagga taggtataca tgttgatgcg ggttttactg
     1561 atgcatatac agagatgctt tttgttcgct tggttgtgat gatgtggtgt ggttgggcgg
     1621 tcgttcattc gttctagatc ggagtagaat actgtttcaa actacctggt gtatttatta
     1681 attttggaac tgtatgtgtg tgtcatacat cttcatagtt acgagtttaa gatggatgga
     1741 aatatcgatc taggataggt atacatgttg atgtgggttt tactgatgca tatacatgat
     1801 ggcatatgca gcatctattc atatgctcta accttgagta cctatctatt ataataaaca
     1861 agtatgtttt ataattattt tgatcttgat atacttggat gatggcatat gcagcagcta
     1921 tatgtggatt tttttagccc tgccttcata cgctatttat ttgcttggta ctgtttcttt
     1981 tgtcgatgct caccctgttg tttggtgtta cttctgcagg aatggagaag aaatactcta
     2041 ttgggctggc cattggcacc aattccgttg gatgggccgt gatcactgat gattataagg
     2101 tcccctccaa gaagttcaag gttctgggta ataccaacag gaagagcata aaaaagaatc
     2161 ttatgggcgc tctgctcttt gatagcggag aaacggcgga agctacacgg ctcaagcgga
     2221 cagcgaggcg ccgctatacc aggcggaaga accggattag ataccttcag gaaatcttcg
     2281 cgaacgagat ggccaagctc gatgactcat tcttccagcg ccttgaggaa tcgttcctcg
     2341 ttgaggagga caaaaagaac gagcgtcacc ccatctttgg taatctcgcg gacgaggttg
     2401 cataccacag gaattatccg accatctacc acctgaggaa gaagttggcg gacagcccgg
     2461 agaaggcaga cctgcggctc atctacctcg cgctggcgca catcatcaaa ttccgggggc
     2521 attttttgat cgaagggaaa ctcaatgctg agaactcgga cgtggcaaag ctcttctatc
     2581 agcttatcca aacctataat cagctgttcg aggaatctcc gttggatgag atagaggttg
     2641 acgctaaggg tatcctctcc gctcgtcttt cgaagtccaa acgcctggag aaactgatcg
     2701 ccgttttccc gaacgagaag aagaacgggc tcttcggaaa cataatcgcg cttgccctgg
     2761 gactgacgcc caacttcaag tccaacttcg acttgacgga ggatgccaaa cttcagcttt
     2821 caaaggacac atatgacgat gatttggacg agttgctcgg ccaaattggc gaccaatacg
     2881 ctgacctgtt ctctgccgca aaaaacctgt ccgatgctat tctgctgagc gatattctcc
     2941 ggagcaactc cgaggtgact aaggcaccgt tgtccgccag catggtgaaa cgctacgacg
     3001 agcaccacca ggacctggct ctcctcaaga ccttggttag gcaacagttt cccgagaagt
     3061 acgccgaaat ctttaaggat gacaccaaga acggctatgc cggttacgtt ggcatcggaa
     3121 tcaagcatag gaagaggacc actaaactcg ccacacagga ggaattctat aaatttatca
     3181 agcctattct ggagaagatg gacggagctg aggagctgct cgccaagctg aacagggacg
     3241 atctcctgag gaagcagcgg acctttgata atggctcgat tcctcatcaa atccatctga
     3301 aggagcttca tgccatactg cgcaggcaag aagagtttta cccatttctg aaggagaatc
     3361 gggagaagat cgaaaaaatc ctgacattcc gcatacccta ctacgtgggc ccgcttgccc
     3421 ggggaaactc tcgttttgct tggctcacga ggaagagcga ggaggcgata acgccatgga
     3481 acttcgagga ggtcgtggat aaaggcgcca gcgcgcaatc gtttattgag cgtatgacaa
     3541 acttcgacga gcagctccca aacaagaagg tgttgccaaa gcacagcctc ctgtacgagt
     3601 acttcacggt ttacaatgag ctcaccaaag tcaagtacgt tactgagcgt atgcgtaagc
     3661 cggagttcct ctctggggag cagaagaagg caatcgttga cctgctcttc aaaacgaacc
     3721 gtaaggtgac cgtcaagcag ctgaaggaag attacttcaa aaagatcgaa tgttttgact
     3781 ccgtggagat tataggagtt gaggataggt ttaatgcctc actcggcacc taccatgact
     3841 tgctcaagat catcaaagac aaggatttcc tggacaatga agagaacgag gatatccttg
     3901 aggacatcgt gctcacactc accctcttcg aggaccggga gatgatcgag gagcgcctca
     3961 agacatacgc ccacctcttt gacgacaagg tgatgaaaca actcaagcgc aggcactaca
     4021 cgggttgggg acgcctctcc cgcaagatga ttaatggtat cagggacaaa cagtccggta
     4081 agacgattct cgatttcttg aagtccgatg gtttctctaa tcgcaacttc atgcagctga
     4141 tccacgacga ctcactcacc ttcaaggagg agattgaaaa ggctcaagtg tccgggcagg
     4201 gagatagcct ccacgagcaa atcgcagatc tcgccggatc gcctgcaatt aagaagggca
     4261 ttcttcaaac tgtcaaaatc gtcgacgagc tggtcaaggt tatgggccac aagccagaaa
     4321 acatcgttat cgagatggct agggagaacc agaccactac caagggactc caacagtccc
     4381 gggagcggaa gaagaggatt gaggagggga tcaaggaact cgagtctcaa attctcaagg
     4441 agaatccggt ggagaacact caactccaga acgagaagct ctacctgtac tacctgcaga
     4501 acggcaggga tatgtatgtc gatcaggagc tggacatcaa caggctctcc gactacgacg
     4561 tcgaccatat cgtcccgcaa tctttcatta aggacgactc aatcgataac aaggtcctca
     4621 cgcgctccgt cgagaacaga ggtaagtcag acaacgtgcc gtcggaggag gtggtgaaga
     4681 agatgaaaaa ctactggcgt caactcctca acgctaaatt gattacgcag agaaaattcg
     4741 ataacttgac caaagcggaa cgcggcggtc tgtcggaggc cgacaaagcc ggcttcatta
     4801 agcggcaact cgtggagaca cgccagatca caaagcacgt cgccaggatt cttgactcgc
     4861 ggatgaatac taaacgcgac aagaacgata agccaatcag agaagtcaag gtgattacac
     4921 tgaagagcaa actggtcagc gacttccgca aggatttcca gctttataag gtgagggaca
     4981 taaacaatta tcaccacgcc cacgacgcgt acctcaacgc ggtggttggc accgcactca
     5041 ttaagaaata cccgaagctc gagtcagagt tcgtctacgg cgactataaa gtgtacgacg
     5101 ttcggaaaat gatagcaaag tccgagcaag agatcggcaa ggccacggcc aagagattct
     5161 tctactccaa tattatgaat ttctttaaga cggaggttaa gttggccaac ggagagatcc
     5221 ggaagcggcc tctcattgaa actaatggcg agacaggcga ggtggtctgg aacaaggaaa
     5281 aggactttgc taccgtccgc aaggtcctcg cgatgccgca agtgaacatt gtcaaaaaga
     5341 cggaagttca gacaggcggg ttctcgaagg agtcaatctt gtcgaagcgt gagtccgcca
     5401 agctcatccc acgcaagaag gggtgggata cgcggaagta tggaggtttc gggtccccca
     5461 ctgttgcata ctcaattctc gtggtcgcca aggtggagaa gggaaaagcc aagaagctca
     5521 aaagcgtgaa ggtcctcgtg ggtataacga taatggaaaa gggtagctac gagaaggacc
     5581 caatcggttt cctcgaagcc aaggggtaca aagatatcaa aaaggagctt atattcaaac
     5641 tgccgaagta ctcattgttt gagctggaga acggcaggcg gcggatgttg gcctcggcga
     5701 cagaactgca aaaggcgaac gagctcgtct tgcctcagca tctcgttagg cttctctact
     5761 acacccaaaa tatctctgcg actacagggt ctaacaacct cggttacatc gagcagcata
     5821 gggaggaatt caaggagatc tttgagaaga tcatcgactt ctccgagaag tacatcctta
     5881 agaacaaagt gaactccaac ttgaagtcga gcttcgacga gcagttcgct gtttccgatt
     5941 ctattctcct gtcgaactca ttcgtgtcac ttctgaagta cacgtcgttc ggcgcatctg
     6001 gcggctttac cttcctcgac ctcgacgtga agcaaggccg gctccgctac caaaccgtga
     6061 ccgaggttct cgacgcgaca ttgatctacc agtccattac cggcctgtac gaaacgagga
     6121 cggacctctc acagctcgga ggggactcta gagcggaccc aaagaagaag cgcaaggtgt
     6181 aggcttgtca agcagatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg
     6241 ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa
     6301 ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat
     6361 tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc
     6421 gcgcggtgtc atctatgtta ctagatcgac gctactattg tcttctgcac gaagtggttt
     6481 aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat
     6541 tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt
     6601 gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc agccggaact
     6661 gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca ggcgtgttcc
     6721 accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc
     6781 acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag cgggtacggc
     6841 tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac
     6901 ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg
     6961 atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc
     7021 agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt
     7081 aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga ccagcccagg
     7141 tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg
     7201 tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc gacgccggtg
     7261 taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc
     7321 gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct
     7381 cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc
     7441 tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc
     7501 tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac
     7561 ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc ggccgatggc
     7621 gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc
     7681 tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt
     7741 gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc
     7801 ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac tcacgccggg
     7861 gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag ataatccacc
     7921 ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta cttggtattc
     7981 cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc gtgggcctcg
     8041 gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt gtcgccggca
     8101 tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt
     8161 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     8221 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     8281 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8341 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8401 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8461 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8521 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8581 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8641 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     8701 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8761 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8821 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8881 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8941 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     9001 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     9061 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     9121 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     9181 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     9241 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     9301 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     9361 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9421 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9481 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9541 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9601 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9661 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9721 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9781 tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg agcggataca
     9841 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat tggccagcgc
     9901 tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag
     9961 gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg
    10021 gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag
    10081 caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg
    10141 attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac
    10201 tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt
    10261 gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt
    10321 aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc
    10381 gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt
    10441 ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc
    10501 cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg
    10561 ccgccagccc agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag atcctgatct
    10621 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
    10681 ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa
    10741 taacacattg cggacgtttt taatgtactg gggtggatgc agtgggcccc ac
//
