LOCUS       sequence-100122-        2909 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Vector for conventional cloning that contains AtU6-26 promoter and 
            sgRNA backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2909)
  AUTHORS   Fauser F, Schiml S, Puchta H
  TITLE     Both CRISPR/Cas-based nucleases and nickases can be used efficiently
            for genome engineering in Arabidopsis thaliana.
  JOURNAL   Plant J. 2014 Jul;79(2):348-59. doi: 10.1111/tpj.12554. Epub 2014 
            Jun 17.
  PUBMED    24836556
REFERENCE   2  (bases 1 to 2909)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1111/tpj.12554";
            journalName: "Plant J"; date: "2014-07"; volume: "79"; issue: "2"; 
            pages: "348-59"
COMMENT     Sequence Label: pChimera
FEATURES             Location/Qualifiers
     source          1..2909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        798..873
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(885..909)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1052..1069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1162..1750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1242..1261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1921..2712,2713..2781))
                     /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     2544..2563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2782..2886)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
      361 aggccacgtg tcttgtccag agctccctag gctttttttc ttcttcttcg ttcatacagt
      421 ttttttttgt ttatcagctt acattttctt gaaccgtagc tttcgttttc ttctttttaa
      481 ctttccattc ggagtttttg tatcttgttt catagtttgt cccaggatta gaatgattag
      541 gcatcgaacc ttcaagaatt tgattgaata aaacatcttc attcttaaga tatgaagata
      601 atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta tatgggaaag
      661 aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa aagtcccaca
      721 tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag tagtgattgg
      781 ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      841 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctagacccag ctttcttgta
      901 caaagttggc attacctagg ggtacctgga gcacaagact ggcctcatgg gccttccgct
      961 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct
     1021 gtttccttgc gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg
     1081 ttcgggtaaa gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa
     1141 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1201 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1261 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1321 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1381 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1441 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1501 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1561 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1621 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1681 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1741 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1801 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1861 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1921 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1981 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2041 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     2101 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2161 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2221 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2281 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2341 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2401 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2461 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2521 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     2581 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2641 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2701 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2761 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2821 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2881 tccgcgcaca tttccccgaa aagtgccac
//
