LOCUS       sequence-257172-        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     complement(31..52)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     243..262
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     476..498
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     490..507
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     491..507
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     514..533
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        514..532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     557..573
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     558..574
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2135..2153)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2174..2190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2174..2190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2187..2209)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2198..2214
                     /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(2222..2228,2229..2246,2247..2252))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2267..2288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2405..2422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2576..3164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2656..2675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3335..4126,4127..4195))
                     /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     3958..3977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4196..4300)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4326..4432,1..349))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
       61 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      121 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      181 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      241 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
      301 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
      361 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      421 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      481 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
      541 ggtaccgggc cccccctcga ggtcgacggt atcgataagc tttgagagta caatgatgaa
      601 cctagattaa tcaatgccaa agtctgaaaa atgcaccctc agtctatgat ccagaaaatc
      661 aagattgctt gaggccctgt tcggttgttc cggattagag ccccggatta attcctagcc
      721 ggattacttc tctaatttat atagattttg atgagctgga atgaatcctg gcttattccg
      781 gtacaaccga acaggccctg aaggatacca gtaatcgctg agctaaattg gcatgctgtc
      841 agagtgtcag tattgcagca aggtagtgag ataaccggca tcatggtgcc agtttgatgg
      901 caccattagg gttagagatg gtggccatgg gcgcatgtcc tggccaactt tgtatgatat
      961 atggcagggt gaataggaaa gtaaaattgt attgtaaaaa gggatttctt ctgtttgtta
     1021 gcgcatgtac aaggaatgca agttttgagc gagggggcat caaagatctg gctgtgtttc
     1081 cagctgtttt tgttagcccc atcgaatcct tgacataatg atcccgctta aataagcaac
     1141 ctcgcttgta tagttccttg tgctctaaca cacgatgatg ataagtcgta aaatagtggt
     1201 gtccaaagaa tttccaggcc cagttgtaaa agctaaaatg ctattcgaat ttctactagc
     1261 agtaagtcgt gtttagaaat tattttttta tatacctttt ttccttctat gtacagtagg
     1321 acacagtgtc agcgccgcgt tgacggagaa tatttgcaaa aaagtaaaag agaaagtcat
     1381 agcggcgtat gtgccaaaaa cttcgtcaca gagagggcca taagaaacat ggcccacggc
     1441 ccaatacgaa gcaccgcgac gaagcccaaa cagcagtccg taggtggagc aaagcgctgg
     1501 gtaatacgca aacgttttgt cccaccttga ctaatcacaa gagtggagcg taccttataa
     1561 accgagccgc aagcaccgaa ttgtttttcc cgcacacgcg ggggtgatcc ttctaaacgc
     1621 taaagaggaa gaggacagaa agagtttttc ccgcacacgc gggggtgatc cttttttttg
     1681 cggccgcgaa ttcctgcagg gccctcttgt cggaccagtt gcccaccacg ttggtgagct
     1741 cggtgaggcc cttcattgag aggaaggagg tcatgaggtg cctaccgatg tgggacttgg
     1801 ggccgttctt gatggcgaag atggagtagg gggcgttctt cttgagggcc ttgttgtagg
     1861 acctcacgag gttgtccttg aggagctggt actcctgctt gttggaggag gagttgccgg
     1921 tcctgttcac cctcttgagc acgggctctg agttcctgag gaactcgtcg aggtacacga
     1981 gggggtcgat cctgccgcga gcggagaaga agtagatgtg cctggacacg gaggtcttgg
     2041 tctcggtcac gaggcactgg atgatcacgc cgaggtactt gttctgacta gttctagagc
     2101 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta atttcgagct
     2161 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2221 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2281 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2341 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2401 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2461 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2521 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2761 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3001 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3181 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3241 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3301 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3361 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3421 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3481 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3541 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3601 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3661 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3721 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3781 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3841 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3901 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3961 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4021 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4081 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4141 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4201 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4261 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4321 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4381 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aa
//
