LOCUS       sequence-221917-        4880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Also known as LBJJ429. PCR template for sgRNA-EF with polyT, 67bp or
            192bp terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4880)
  AUTHORS   Castel B, Tomlinson L, Locci F, Yang Y, Jones JDG
  TITLE     Optimization of T-DNA architecture for Cas9-mediated mutagenesis in 
            Arabidopsis.
  JOURNAL   PLoS One. 2019 Jan 9;14(1):e0204778. doi: 
            10.1371/journal.pone.0204778. eCollection 2019.
  PUBMED    30625150
REFERENCE   2  (bases 1 to 4880)
  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: "PLoS One.";
            date: "2019-01-9"; volume: "14(1):e0204778. doi"; pages: " 
            10.1371/journal.pone.0204778. eCollection 2019"
COMMENT     Sequence Label: LBJJ382
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    583..607
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(763..1911)
                     /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(2247..2835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2327..2346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3006..3797,3798..3866))
                     /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     3629..3648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3867..3971)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4058..4689
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4781..4805
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caaactagaa ttcgagctcg gagcatcttc attcttaaga tatgaagata
       61 atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta tatgggaaag
      121 aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa aagtcccaca
      181 tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag tagtgattgc
      241 gtatcttcgg ccatgaagcg tttaagagct atgctggaaa cagcatagca agtttaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tttgcaaaat
      361 tttccagatc gatttcttct tcctctgttc ttcggcgttc aatttctggg gttttctctt
      421 cgttttctgt aactgaaacc taaaatttga cctaaaaaaa atctcaaata atatgattca
      481 gtggttttgt acttttcagt tagttgagtt ttgcagttcc gatgagataa accaatacgc
      541 tttacttgtc ttctgcacga agtggtttaa actatcagtg tttgacagga tatattggcg
      601 ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa
      661 aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc cagatcaggc
      721 gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac
      781 caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc
      841 ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg
      901 gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg
      961 tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc
     1021 agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc
     1081 acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc
     1141 ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta
     1201 ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat
     1261 cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc
     1321 atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa
     1381 aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc
     1441 agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa
     1501 caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt
     1561 ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat
     1621 agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcaagacg
     1681 acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct
     1741 gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc
     1801 gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc
     1861 cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc
     1921 ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc
     1981 tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt
     2041 ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagcgaccc
     2101 cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga tgcggaagaa
     2161 gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc tgccaggaac
     2221 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2281 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2341 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2401 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2461 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2521 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2581 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2641 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2701 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2761 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2821 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2881 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2941 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3001 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3061 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3121 ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca
     3181 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3241 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3301 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3361 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3421 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3481 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3541 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3601 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3661 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3721 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3781 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3841 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3901 caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3961 ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg
     4021 aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa
     4081 gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa
     4141 acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa
     4201 aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc
     4261 aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct
     4321 catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt
     4381 gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct
     4441 ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg
     4501 ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt
     4561 tccgcgaggt atccacaacg ccggcggccg cggtgtctcg cacacggctt cgacggcgtt
     4621 tctggcgcgt ttgcagggcc atagacggcc gccagcccag cggcgagggc aaccagcccg
     4681 gtgagcgtcg caaaggagat cctgatctga ctgatgggct gcctgtatcg agtggtgatt
     4741 ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg
     4801 taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg
     4861 gtggatgcag tgggccccac
//
