LOCUS       sequence-254141-        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     63..85
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     77..94
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     78..94
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     101..120
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        101..119
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     144..160
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     145..161
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1722..1740)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1761..1777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1761..1777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1774..1796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1785..1801
                     /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(1809..1815,1816..1833,1834..1839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1854..1875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1992..2009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2163..2751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2243..2262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2922..3713,3714..3782))
                     /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     3545..3564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3783..3887)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3913..4368)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4050..4071)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4262..4281
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
       61 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
      121 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttt gagagtacaa
      181 tgatgaacct agattaatca atgccaaagt ctgaaaaatg caccctcagt ctatgatcca
      241 gaaaatcaag attgcttgag gccctgttcg gttgttccgg attagagccc cggattaatt
      301 cctagccgga ttacttctct aatttatata gattttgatg agctggaatg aatcctggct
      361 tattccggta caaccgaaca ggccctgaag gataccagta atcgctgagc taaattggca
      421 tgctgtcaga gtgtcagtat tgcagcaagg tagtgagata accggcatca tggtgccagt
      481 ttgatggcac cattagggtt agagatggtg gccatgggcg catgtcctgg ccaactttgt
      541 atgatatatg gcagggtgaa taggaaagta aaattgtatt gtaaaaaggg atttcttctg
      601 tttgttagcg catgtacaag gaatgcaagt tttgagcgag ggggcatcaa agatctggct
      661 gtgtttccag ctgtttttgt tagccccatc gaatccttga cataatgatc ccgcttaaat
      721 aagcaacctc gcttgtatag ttccttgtgc tctaacacac gatgatgata agtcgtaaaa
      781 tagtggtgtc caaagaattt ccaggcccag ttgtaaaagc taaaatgcta ttcgaatttc
      841 tactagcagt aagtcgtgtt tagaaattat ttttttatat accttttttc cttctatgta
      901 cagtaggaca cagtgtcagc gccgcgttga cggagaatat ttgcaaaaaa gtaaaagaga
      961 aagtcatagc ggcgtatgtg ccaaaaactt cgtcacagag agggccataa gaaacatggc
     1021 ccacggccca atacgaagca ccgcgacgaa gcccaaacag cagtccgtag gtggagcaaa
     1081 gcgctgggta atacgcaaac gttttgtccc accttgacta atcacaagag tggagcgtac
     1141 cttataaacc gagccgcaag caccgaattg tttttcccgc acacgcgggg gtgatccttt
     1201 atggacagag ctccaagtga ccgagcaaga gtttttcccg cacacgcggg ggtgatcctt
     1261 ttttttgcgg ccgcgaattc ctgcagggcc ctcttgtcgg accagttgcc caccacgttg
     1321 gtgagctcgg tgaggccctt cattgagagg aaggaggtca tgaggtgcct accgatgtgg
     1381 gacttggggc cgttcttgat ggcgaagatg gagtaggggg cgttcttctt gagggccttg
     1441 ttgtaggacc tcacgaggtt gtccttgagg agctggtact cctgcttgtt ggaggaggag
     1501 ttgccggtcc tgttcaccct cttgagcacg ggctctgagt tcctgaggaa ctcgtcgagg
     1561 tacacgaggg ggtcgatcct gccgcgagcg gagaagaagt agatgtgcct ggacacggag
     1621 gtcttggtct cggtcacgag gcactggatg atcacgccga ggtacttgtt ctgactagtt
     1681 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     1741 tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1801 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1861 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1921 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1981 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2581 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2941 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3001 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3061 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3121 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3181 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3241 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3301 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3361 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3421 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3481 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3541 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3601 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3661 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3721 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3781 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3841 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3901 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3961 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4021 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4081 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4141 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4201 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4261 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4321 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     4381 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta tt
//
