LOCUS       sequence-254132-        4438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 DsRed targeting sgRNA expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4438)
  AUTHORS   Young JK, Gasior SL, Jones S, Wang L, Navarro P, Vickroy B, 
            Barrangou R
  TITLE     The repurposing of type I-E CRISPR-Cascade for gene activation in 
            plants.
  JOURNAL   Commun Biol. 2019 Oct 18;2:383. doi: 10.1038/s42003-019-0637-6. 
            eCollection 2019.
  PUBMED    31646186
REFERENCE   2  (bases 1 to 4438)
  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: "Commun 
            Biol."; date: "2019-10-18"; volume: "2:383. doi"; pages: " 
            10.1038/s42003-019-0637-6. eCollection 2019"
COMMENT     Sequence Label: PV384
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..77
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     69..86
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     70..86
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     93..112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        93..111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     136..152
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     137..153
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        1180..1255
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(1720..1738)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1759..1775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1759..1775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1772..1794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1783..1799
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1807..1813,1814..1831,1832..1837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1852..1873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1990..2007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2161..2749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2241..2260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2920..3711,3712..3780))
                     /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     3543..3562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3781..3885)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3911..4366)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4048..4069)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4260..4279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
       61 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
      121 gtaccgggcc ccccctcgag gtcgacggta tcgataagct ttgagagtac aatgatgaac
      181 ctagattaat caatgccaaa gtctgaaaaa tgcaccctca gtctatgatc cagaaaatca
      241 agattgcttg aggccctgtt cggttgttcc ggattagagc cccggattaa ttcctagccg
      301 gattacttct ctaatttata tagattttga tgagctggaa tgaatcctgg cttattccgg
      361 tacaaccgaa caggccctga aggataccag taatcgctga gctaaattgg catgctgtca
      421 gagtgtcagt attgcagcaa ggtagtgaga taaccggcat catggtgcca gtttgatggc
      481 accattaggg ttagagatgg tggccatggg cgcatgtcct ggccaacttt gtatgatata
      541 tggcagggtg aataggaaag taaaattgta ttgtaaaaag ggatttcttc tgtttgttag
      601 cgcatgtaca aggaatgcaa gttttgagcg agggggcatc aaagatctgg ctgtgtttcc
      661 agctgttttt gttagcccca tcgaatcctt gacataatga tcccgcttaa ataagcaacc
      721 tcgcttgtat agttccttgt gctctaacac acgatgatga taagtcgtaa aatagtggtg
      781 tccaaagaat ttccaggccc agttgtaaaa gctaaaatgc tattcgaatt tctactagca
      841 gtaagtcgtg tttagaaatt atttttttat ataccttttt tccttctatg tacagtagga
      901 cacagtgtca gcgccgcgtt gacggagaat atttgcaaaa aagtaaaaga gaaagtcata
      961 gcggcgtatg tgccaaaaac ttcgtcacag agagggccat aagaaacatg gcccacggcc
     1021 caatacgaag caccgcgacg aagcccaaac agcagtccgt aggtggagca aagcgctggg
     1081 taatacgcaa acgttttgtc ccaccttgac taatcacaag agtggagcgt accttataaa
     1141 ccgagccgca agcaccgaat tgaggacaga aagaaaatcg ttttagagct agaaatagca
     1201 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     1261 ttttgcggcc gcgaattcct gcagggccct cttgtcggac cagttgccca ccacgttggt
     1321 gagctcggtg aggcccttca ttgagaggaa ggaggtcatg aggtgcctac cgatgtggga
     1381 cttggggccg ttcttgatgg cgaagatgga gtagggggcg ttcttcttga gggccttgtt
     1441 gtaggacctc acgaggttgt ccttgaggag ctggtactcc tgcttgttgg aggaggagtt
     1501 gccggtcctg ttcaccctct tgagcacggg ctctgagttc ctgaggaact cgtcgaggta
     1561 cacgaggggg tcgatcctgc cgcgagcgga gaagaagtag atgtgcctgg acacggaggt
     1621 cttggtctcg gtcacgaggc actggatgat cacgccgagg tacttgttct gactagttct
     1681 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaatttc
     1741 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1801 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1861 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1921 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1981 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2041 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2101 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2161 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2221 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2581 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2701 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2761 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2821 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2881 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2941 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3001 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3061 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3121 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3181 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3241 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3301 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3361 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3421 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3481 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3541 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3601 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3661 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3721 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3781 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3841 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3901 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3961 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4021 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4081 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4141 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4201 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4261 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4321 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     4381 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagc
//
