LOCUS       sequence-254133-        4432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SthCascade DsRed targeting crRNA expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4432)
  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 4432)
  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: PV422
FEATURES             Location/Qualifiers
     source          1..4432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     72..89
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     73..89
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     96..115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        96..114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     139..155
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     140..156
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1717..1735)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1756..1772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1756..1772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1769..1791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1780..1796
                     /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(1804..1810,1811..1828,1829..1834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1849..1870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1987..2004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2158..2746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2238..2257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2917..3708,3709..3777))
                     /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     3540..3559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3778..3882)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3908..4363)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4045..4066)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4257..4276
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
       61 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
      121 tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gctttgagag tacaatgatg
      181 aacctagatt aatcaatgcc aaagtctgaa aaatgcaccc tcagtctatg atccagaaaa
      241 tcaagattgc ttgaggccct gttcggttgt tccggattag agccccggat taattcctag
      301 ccggattact tctctaattt atatagattt tgatgagctg gaatgaatcc tggcttattc
      361 cggtacaacc gaacaggccc tgaaggatac cagtaatcgc tgagctaaat tggcatgctg
      421 tcagagtgtc agtattgcag caaggtagtg agataaccgg catcatggtg ccagtttgat
      481 ggcaccatta gggttagaga tggtggccat gggcgcatgt cctggccaac tttgtatgat
      541 atatggcagg gtgaatagga aagtaaaatt gtattgtaaa aagggatttc ttctgtttgt
      601 tagcgcatgt acaaggaatg caagttttga gcgagggggc atcaaagatc tggctgtgtt
      661 tccagctgtt tttgttagcc ccatcgaatc cttgacataa tgatcccgct taaataagca
      721 acctcgcttg tatagttcct tgtgctctaa cacacgatga tgataagtcg taaaatagtg
      781 gtgtccaaag aatttccagg cccagttgta aaagctaaaa tgctattcga atttctacta
      841 gcagtaagtc gtgtttagaa attatttttt tatatacctt ttttccttct atgtacagta
      901 ggacacagtg tcagcgccgc gttgacggag aatatttgca aaaaagtaaa agagaaagtc
      961 atagcggcgt atgtgccaaa aacttcgtca cagagagggc cataagaaac atggcccacg
     1021 gcccaatacg aagcaccgcg acgaagccca aacagcagtc cgtaggtgga gcaaagcgct
     1081 gggtaatacg caaacgtttt gtcccacctt gactaatcac aagagtggag cgtaccttat
     1141 aaaccgagcc gcaagcaccg aattgttttt cccgcacacg cgggggtgat ccttctaaac
     1201 gctaaagagg aagaggacag aaagagtttt tcccgcacac gcgggggtga tccttttttt
     1261 tgcggccgcg aattcctgca gggccctctt gtcggaccag ttgcccacca cgttggtgag
     1321 ctcggtgagg cccttcattg agaggaagga ggtcatgagg tgcctaccga tgtgggactt
     1381 ggggccgttc ttgatggcga agatggagta gggggcgttc ttcttgaggg ccttgttgta
     1441 ggacctcacg aggttgtcct tgaggagctg gtactcctgc ttgttggagg aggagttgcc
     1501 ggtcctgttc accctcttga gcacgggctc tgagttcctg aggaactcgt cgaggtacac
     1561 gagggggtcg atcctgccgc gagcggagaa gaagtagatg tgcctggaca cggaggtctt
     1621 ggtctcggtc acgaggcact ggatgatcac gccgaggtac ttgttctgac tagttctaga
     1681 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag
     1741 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1801 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1861 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2581 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3001 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3061 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3121 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3181 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3241 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3301 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3361 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3421 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3481 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3541 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3601 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3661 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3721 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3781 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3841 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3901 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3961 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4021 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4081 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4141 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4201 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4261 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4321 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     4381 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg cc
//
