LOCUS       sequence-254114-        6334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CasE constitutive expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6334)
  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 6334)
  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: PV246
FEATURES             Location/Qualifiers
     source          1..6334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..39
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     31..48
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     32..48
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     55..74
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        55..73
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        108..2103
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             2770..2790
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(3179..3197)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3180..3197)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3215..3231)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3215..3231)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3228..3250)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3239..3255
                     /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)."
     protein_bind    3315..3336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3453..3470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3624..4212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3704..3723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4386..4405)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4397..5212
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4397..5212
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4470..4489)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5506..5524
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5562..5584)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5684..5703
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5770..6225)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5907..5928)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6119..6138
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
       61 gactcactat agggcgaatt gggttacccg gaccgaagct tgcatgcctg cagtgcagcg
      121 tgacccggtc gtgcccctct ctagagataa tgagcattgc atgtctaagt tataaaaaat
      181 taccacatat tttttttgtc acacttgttt gaagtgcagt ttatctatct ttatacatat
      241 atttaaactt tactctacga ataatataat ctatagtact acaataatat cagtgtttta
      301 gagaatcata taaatgaaca gttagacatg gtctaaagga caattgagta ttttgacaac
      361 aggactctac agttttatct ttttagtgtg catgtgttct cctttttttt tgcaaatagc
      421 ttcacctata taatacttca tccattttat tagtacatcc atttagggtt tagggttaat
      481 ggtttttata gactaatttt tttagtacat ctattttatt ctattttagc ctctaaatta
      541 agaaaactaa aactctattt tagttttttt atttaataat ttagatataa aatagaataa
      601 aataaagtga ctaaaaatta aacaaatacc ctttaagaaa ttaaaaaaac taaggaaaca
      661 tttttcttgt ttcgagtaga taatgccagc ctgttaaacg ccgtcgacga gtctaacgga
      721 caccaaccag cgaaccagca gcgtcgcgtc gggccaagcg aagcagacgg cacggcatct
      781 ctgtcgctgc ctctggaccc ctctcgagag ttccgctcca ccgttggact tgctccgctg
      841 tcggcatcca gaaattgcgt ggcggagcgg cagacgtgag ccggcacggc aggcggcctc
      901 ctcctcctct cacggcaccg gcagctacgg gggattcctt tcccaccgct ccttcgcttt
      961 cccttcctcg cccgccgtaa taaatagaca ccccctccac accctctttc cccaacctcg
     1021 tgttgttcgg agcgcacaca cacacaacca gatctccccc aaatccaccc gtcggcacct
     1081 ccgcttcaag gtacgccgct cgtcctcccc cccccccctc tctaccttct ctagatcggc
     1141 gttccggtcc atgcatggtt agggcccggt agttctactt ctgttcatgt ttgtgttaga
     1201 tccgtgtttg tgttagatcc gtgctgctag cgttcgtaca cggatgcgac ctgtacgtca
     1261 gacacgttct gattgctaac ttgccagtgt ttctctttgg ggaatcctgg gatggctcta
     1321 gccgttccgc agacgggatc gatttcatga ttttttttgt ttcgttgcat agggtttggt
     1381 ttgccctttt cctttatttc aatatatgcc gtgcacttgt ttgtcgggtc atcttttcat
     1441 gctttttttt gtcttggttg tgatgatgtg gtctggttgg gcggtcgttc tagatcggag
     1501 tagaattctg tttcaaacta cctggtggat ttattaattt tggatctgta tgtgtgtgcc
     1561 atacatattc atagttacga attgaagatg atggatggaa atatcgatct aggataggta
     1621 tacatgttga tgcgggtttt actgatgcat atacagagat gctttttgtt cgcttggttg
     1681 tgatgatgtg gtgtggttgg gcggtcgttc attcgttcta gatcggagta gaatactgtt
     1741 tcaaactacc tggtgtattt attaattttg gaactgtatg tgtgtgtcat acatcttcat
     1801 agttacgagt ttaagatgga tggaaatatc gatctaggat aggtatacat gttgatgtgg
     1861 gttttactga tgcatataca tgatggcata tgcagcatct attcatatgc tctaaccttg
     1921 agtacctatc tattataata aacaagtatg ttttataatt attttgatct tgatatactt
     1981 ggatgatggc atatgcagca gctatatgtg gattttttta gccctgcctt catacgctat
     2041 ttatttgctt ggtactgttt cttttgtcga tgctcaccct gttgtttggt gttacttctg
     2101 caggtcgact ctagaggatc catgtacatc agcagggtcg agatcgaccg atataatagg
     2161 cgcaaggtca gggacctcac ccacgtcggc gcctaccacg cctgggtcga ggagagcttc
     2221 cccagcgagc tcgagcaatc aatcaggacc aggaagctct ggcgcatcga ccgcatccag
     2281 ggcaaggact acctcatcat cgtcagcaag gagaagccca acctccagaa gctcgaaaaa
     2341 tatggcgtcg tcggcagcgc ccagaccaag gactaccagc acttcctcga cagcatcaag
     2401 accggcttca ggatgaactt caggatcgtc ctcaaccccg tcatcagcat caccagcccc
     2461 gacaaatcaa aacgcggcgt cgtcaagccc cacgtcacca ttaaacacca gatggacttc
     2521 ctcgtcaagc agagcgaaaa aaacggcttc agcctcgtcg gcgagagctt ctccatcgtc
     2581 gagaggggct acgaggtgtt ccagaagccc gagaagccca tcaggctcag caaggtcgtc
     2641 tacgagggca ccctcaccgt cagcgacgac acgctcttca agaagatgct caccgagggc
     2701 gtcggcaaga agaaagcata tggcttcggc ctcatgaccg tcgtccccct cgaggagagc
     2761 agggccgacc ccaagaagaa gaggaaggtc tagtcttcgt aagttaacct agacttgtcc
     2821 atcttctgga ttggccaact taattaatgt atgaaataaa aggatgcaca catagtgaca
     2881 tgctaatcac tataatgtgg gcatcaaagt tgtgtgttat gtgtaattac tagttatctg
     2941 aataaaagag aaagagatca tccatatttc ttatcctaaa tgaatgtcac gtgtctttat
     3001 aattctttga tgaaccagat gcatttcatt aaccaaatcc atatacatat aaatattaat
     3061 catatataat taatatcaat tgggttagca aaacaaatct agtctaggtg tgttttgcga
     3121 atgcggccgc caccgcggtg gagctcgaat tccggtccgg gtcacccagc ttgagtattc
     3181 tatagtgtca cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3241 gttatccgct cacaattcca cacaacatac gagccggaag cataaagcat aaagtgtaaa
     3301 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3361 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3421 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3481 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3541 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3601 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3661 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3721 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3781 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3841 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3901 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3961 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4021 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4081 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4141 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4201 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4261 aactcacgtt aagggatttt ggtcatggag ccacgttgtg tctcaaaatc tctgatgtta
     4321 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     4381 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcga ggccgcgatt
     4441 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     4501 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     4561 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     4621 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     4681 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     4741 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     4801 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     4861 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     4921 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     4981 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     5041 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     5101 cctcggtgag ttttctcctt cattacagaa acggcttttt cagaaatatg gtattgataa
     5161 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     5221 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcggctttgt
     5281 tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacaacgca
     5341 gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta caacaaagct
     5401 ctcatcaacc gtggctccct cactttctgg ctggatgatg gggcgattca ggcctggtat
     5461 gagtcagcaa caccttcttc acgagccatg acattaacct ataaaaatag gcgtatcacg
     5521 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     5581 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     5641 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     5701 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5761 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5821 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5881 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5941 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     6001 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     6061 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6121 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6181 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     6241 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6301 gaaaggggga tgtgctgcaa ggcgattaag ttgg
//
