LOCUS       sequence-258180-        4432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SthCascade R promoter targeting crRNA3 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: PV569
FEATURES             Location/Qualifiers
     source          1..4432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1578..1596)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1617..1633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1617..1633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1630..1652)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1641..1657
                     /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(1665..1671,1672..1689,1690..1695))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1710..1731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1848..1865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2019..2607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2099..2118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2778..3569,3570..3638))
                     /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     3401..3420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3639..3743)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3769..4224)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3906..3927)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4118..4137
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4351..4373
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4365..4382
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4366..4382
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4389..4408
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4389..4407
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(4432,1..16)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cgaggtcgac ggtatcgata agctttgaga gtacaatgat gaacctagat taatcaatgc
       61 caaagtctga aaaatgcacc ctcagtctat gatccagaaa atcaagattg cttgaggccc
      121 tgttcggttg ttccggatta gagccccgga ttaattccta gccggattac ttctctaatt
      181 tatatagatt ttgatgagct ggaatgaatc ctggcttatt ccggtacaac cgaacaggcc
      241 ctgaaggata ccagtaatcg ctgagctaaa ttggcatgct gtcagagtgt cagtattgca
      301 gcaaggtagt gagataaccg gcatcatggt gccagtttga tggcaccatt agggttagag
      361 atggtggcca tgggcgcatg tcctggccaa ctttgtatga tatatggcag ggtgaatagg
      421 aaagtaaaat tgtattgtaa aaagggattt cttctgtttg ttagcgcatg tacaaggaat
      481 gcaagttttg agcgaggggg catcaaagat ctggctgtgt ttccagctgt ttttgttagc
      541 cccatcgaat ccttgacata atgatcccgc ttaaataagc aacctcgctt gtatagttcc
      601 ttgtgctcta acacacgatg atgataagtc gtaaaatagt ggtgtccaaa gaatttccag
      661 gcccagttgt aaaagctaaa atgctattcg aatttctact agcagtaagt cgtgtttaga
      721 aattattttt ttatatacct tttttccttc tatgtacagt aggacacagt gtcagcgccg
      781 cgttgacgga gaatatttgc aaaaaagtaa aagagaaagt catagcggcg tatgtgccaa
      841 aaacttcgtc acagagaggg ccataagaaa catggcccac ggcccaatac gaagcaccgc
      901 gacgaagccc aaacagcagt ccgtaggtgg agcaaagcgc tgggtaatac gcaaacgttt
      961 tgtcccacct tgactaatca caagagtgga gcgtacctta taaaccgagc cgcaagcacc
     1021 gaattgtttt tcccgcacac gcgggggtga tcctttatgg acagagctcc aagtgaccga
     1081 gcaagagttt ttcccgcaca cgcgggggtg atcctttttt ttgcggccgc gaattcctgc
     1141 agggccctct tgtcggacca gttgcccacc acgttggtga gctcggtgag gcccttcatt
     1201 gagaggaagg aggtcatgag gtgcctaccg atgtgggact tggggccgtt cttgatggcg
     1261 aagatggagt agggggcgtt cttcttgagg gccttgttgt aggacctcac gaggttgtcc
     1321 ttgaggagct ggtactcctg cttgttggag gaggagttgc cggtcctgtt caccctcttg
     1381 agcacgggct ctgagttcct gaggaactcg tcgaggtaca cgagggggtc gatcctgccg
     1441 cgagcggaga agaagtagat gtgcctggac acggaggtct tggtctcggt cacgaggcac
     1501 tggatgatca cgccgaggta cttgttctga ctagttctag agcggccgcc accgcggtgg
     1561 agctccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca
     1621 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1681 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1741 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1801 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1861 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2701 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2761 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2821 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2881 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2941 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3001 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3061 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3121 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3181 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3241 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3301 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3361 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3421 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3481 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3541 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3601 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3661 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3721 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     3781 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3841 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3901 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3961 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4021 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4081 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4141 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4201 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     4261 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4321 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4381 gtgaattgta atacgactca ctatagggcg aattgggtac cgggcccccc ct
//
