LOCUS       sequence-258470-        4432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SthCascade R promoter targeting crRNA1 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: PV564
FEATURES             Location/Qualifiers
     source          1..4432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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)."
     primer_bind     6..28
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     25..41
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     25..41
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        62..80
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1641..1657)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1642..1658)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1682..1701)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1683..1701)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1708..1725)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1708..1724)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1717..1739)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1866..2321
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1953..1972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2163..2184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2347..2451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2452..2520,2521..3312)
                     /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     complement(2670..2689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3483..4071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3972..3991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4225..4242
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4359..4380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4395..4400,4401..4418,4419..4425)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
       61 aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta
      121 gaactagtca gaacaagtac ctcggcgtga tcatccagtg cctcgtgacc gagaccaaga
      181 cctccgtgtc caggcacatc tacttcttct ccgctcgcgg caggatcgac cccctcgtgt
      241 acctcgacga gttcctcagg aactcagagc ccgtgctcaa gagggtgaac aggaccggca
      301 actcctcctc caacaagcag gagtaccagc tcctcaagga caacctcgtg aggtcctaca
      361 acaaggccct caagaagaac gccccctact ccatcttcgc catcaagaac ggccccaagt
      421 cccacatcgg taggcacctc atgacctcct tcctctcaat gaagggcctc accgagctca
      481 ccaacgtggt gggcaactgg tccgacaaga gggccctgca ggaattcgcg gccgcaaaaa
      541 aaaggatcac ccccgcgtgt gcgggaaaaa cgaccgagca agacggtgat ggaagtagac
      601 cctcggatca cccccgcgtg tgcgggaaaa acaattcggt gcttgcggct cggtttataa
      661 ggtacgctcc actcttgtga ttagtcaagg tgggacaaaa cgtttgcgta ttacccagcg
      721 ctttgctcca cctacggact gctgtttggg cttcgtcgcg gtgcttcgta ttgggccgtg
      781 ggccatgttt cttatggccc tctctgtgac gaagtttttg gcacatacgc cgctatgact
      841 ttctctttta cttttttgca aatattctcc gtcaacgcgg cgctgacact gtgtcctact
      901 gtacatagaa ggaaaaaagg tatataaaaa aataatttct aaacacgact tactgctagt
      961 agaaattcga atagcatttt agcttttaca actgggcctg gaaattcttt ggacaccact
     1021 attttacgac ttatcatcat cgtgtgttag agcacaagga actatacaag cgaggttgct
     1081 tatttaagcg ggatcattat gtcaaggatt cgatggggct aacaaaaaca gctggaaaca
     1141 cagccagatc tttgatgccc cctcgctcaa aacttgcatt ccttgtacat gcgctaacaa
     1201 acagaagaaa tcccttttta caatacaatt ttactttcct attcaccctg ccatatatca
     1261 tacaaagttg gccaggacat gcgcccatgg ccaccatctc taaccctaat ggtgccatca
     1321 aactggcacc atgatgccgg ttatctcact accttgctgc aatactgaca ctctgacagc
     1381 atgccaattt agctcagcga ttactggtat ccttcagggc ctgttcggtt gtaccggaat
     1441 aagccaggat tcattccagc tcatcaaaat ctatataaat tagagaagta atccggctag
     1501 gaattaatcc ggggctctaa tccggaacaa ccgaacaggg cctcaagcaa tcttgatttt
     1561 ctggatcata gactgagggt gcatttttca gactttggca ttgattaatc taggttcatc
     1621 attgtactct caaagcttat cgataccgtc gacctcgagg gggggcccgg tacccaattc
     1681 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
     1741 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1801 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1861 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1921 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1981 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2041 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2101 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2161 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2221 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2281 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     2341 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2401 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2461 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2521 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2581 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2641 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2701 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2761 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2821 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2881 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2941 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3001 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3061 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3121 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3181 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3241 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3301 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3361 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3421 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3481 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3541 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3601 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3661 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3721 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3781 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3841 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3901 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3961 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4021 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4081 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4141 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4201 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4261 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     4321 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     4381 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aa
//
