LOCUS       sequence-258471-        6756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CasC constitutive expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6756)
  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 6756)
  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: PV245
FEATURES             Location/Qualifiers
     source          1..6756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..2030
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             3126..3146
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(3535..3553)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3536..3553)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3571..3587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3571..3587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3584..3606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3595..3611
                     /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(3619..3625,3626..3643,3644..3649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3664..3685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3802..3819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3973..4561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4053..4072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4735..4754)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4746..5561
                     /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             4746..5561
                     /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(4819..4838)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5855..5873
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5911..5933)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6033..6052
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6119..6574)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6256..6277)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6468..6487
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6700..6722
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6714..6731
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6715..6731
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6738..6756,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6738..6756
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcgaattggg ttacccggac cgaagcttgc atgcctgcag tgcagcgtga cccggtcgtg
       61 cccctctcta gagataatga gcattgcatg tctaagttat aaaaaattac cacatatttt
      121 ttttgtcaca cttgtttgaa gtgcagttta tctatcttta tacatatatt taaactttac
      181 tctacgaata atataatcta tagtactaca ataatatcag tgttttagag aatcatataa
      241 atgaacagtt agacatggtc taaaggacaa ttgagtattt tgacaacagg actctacagt
      301 tttatctttt tagtgtgcat gtgttctcct ttttttttgc aaatagcttc acctatataa
      361 tacttcatcc attttattag tacatccatt tagggtttag ggttaatggt ttttatagac
      421 taattttttt agtacatcta ttttattcta ttttagcctc taaattaaga aaactaaaac
      481 tctattttag tttttttatt taataattta gatataaaat agaataaaat aaagtgacta
      541 aaaattaaac aaataccctt taagaaatta aaaaaactaa ggaaacattt ttcttgtttc
      601 gagtagataa tgccagcctg ttaaacgccg tcgacgagtc taacggacac caaccagcga
      661 accagcagcg tcgcgtcggg ccaagcgaag cagacggcac ggcatctctg tcgctgcctc
      721 tggacccctc tcgagagttc cgctccaccg ttggacttgc tccgctgtcg gcatccagaa
      781 attgcgtggc ggagcggcag acgtgagccg gcacggcagg cggcctcctc ctcctctcac
      841 ggcaccggca gctacggggg attcctttcc caccgctcct tcgctttccc ttcctcgccc
      901 gccgtaataa atagacaccc cctccacacc ctctttcccc aacctcgtgt tgttcggagc
      961 gcacacacac acaaccagat ctcccccaaa tccacccgtc ggcacctccg cttcaaggta
     1021 cgccgctcgt cctccccccc ccccntctct accttctcta gatcggcgtt ccggtccatg
     1081 catggttagg gcccggtagt tctacttctg ttcatgtttg tgttagatcc gtgtttgtgt
     1141 tagatccgtg ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac acgttctgat
     1201 tgctaacttg ccagtgtttc tctttgggga atcctgggat ggctctagcc gttccgcaga
     1261 cgggatcgat ttcatgattt tttttgtttc gttgcatagg gtttggtttg cccttttcct
     1321 ttatttcaat atatgccgtg cacttgtttg tcgggtcatc ttttcatgct tttttttgtc
     1381 ttggttgtga tgatgtggtc tggttgggcg gtcgttctag atcggagtag aattctgttt
     1441 caaactacct ggtggattta ttaattttgg atctgtatgt gtgtgccata catattcata
     1501 gttacgaatt gaagatgatg gatggaaata tcgatctagg ataggtatac atgttgatgc
     1561 gggttttact gatgcatata cagagatgct ttttgttcgc ttggttgtga tgatgtggtg
     1621 tggttgggcg gtcgttcatt cgttctagat cggagtagaa tactgtttca aactacctgg
     1681 tgtatttatt aattttggaa ctgtatgtgt gtgtcataca tcttcatagt tacgagttta
     1741 agatggatgg aaatatcgat ctaggatagg tatacatgtt gatgtgggtt ttactgatgc
     1801 atatacatga tggcatatgc agcatctatt catatgctct aaccttgagt acctatctat
     1861 tataataaac aagtatgttt tataattatt ttgatcttga tatacttgga tgatggcata
     1921 tgcagcagct atatgtggat ttttttagcc ctgccttcat acgctattta tttgcttggt
     1981 actgtttctt ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag gtcgactcta
     2041 gaggatccat gaccaccgag cagcgcctct tcctcgacat ccacgccatc cagagcctcc
     2101 cccccagcaa catgaacagg gacgacaccg gcagccccaa gaccgcccag tacggcggcg
     2161 tcaggaggag cagagtctcg tcccaagcat ggaagaaggc catgagggag tacttcaaca
     2221 cccacggcga ccactcaaac gtcggcatcc gcaccaagga gatcgtccgc tacgtcgccg
     2281 ataaaatagt cgagctcgac agcagcatca gcatcgagga cgccctcgag aaggcggaca
     2341 aggtcctcat cgccgccgga atcaagaaga agggcgaggt caaggccctc tacttcatgg
     2401 gcgacagcca ggccaagaag ctcgcccagg ccgcctacga caatataaca cataaaaaag
     2461 aactccagaa gctcgcgaac gaggaccccg ccatcgacat cgccctcttc ggcaggatgg
     2521 tcgccgagga ccccaccctg aacgaggact ccagcagcca ggtcgcccac gcgatcagca
     2581 cccaccccgt ccagacggag ttcgacttct tcaccgccgt cgacgacctc gcccccgagg
     2641 acaacgccgg cgccggcatg ctcggcacgg tcgagttcaa cagcagcacg ctctacaggt
     2701 acgccaacgt cgccatccac gagctcagca agcagctcgg caacaaggag agcctcgaga
     2761 acagcctcag cctcttcatc gaggccttcg ccaagagcct ccccaccggc aaggtcaata
     2821 catttgccaa tcaaaccctc ccccagaccc tcatcgtcac cgtgaggagc gacaggcccg
     2881 tcaacctcgt ctccgcgttc gaagagcccg tcaaatcaat agagggctac gtcaccaaga
     2941 gcatcgacaa gctcgccaag gagttcatca aggtcgagaa gttcgtcgac aagcccctca
     3001 tgaccttcta cgtcaccctc cacgagagcg acagcctcaa gcagatcggc gaggagaagg
     3061 acagcgtcag caagctcctc gaggacttca gcgagatgat cgcccagtac atcagccgcg
     3121 ccgaccccaa gaagaagcgc aaggtctagt cttcgtaagt taacctagac ttgtccatct
     3181 tctggattgg ccaacttaat taatgtatga aataaaagga tgcacacata gtgacatgct
     3241 aatcactata atgtgggcat caaagttgtg tgttatgtgt aattactagt tatctgaata
     3301 aaagagaaag agatcatcca tatttcttat cctaaatgaa tgtcacgtgt ctttataatt
     3361 ctttgatgaa ccagatgcat ttcattaacc aaatccatat acatataaat attaatcata
     3421 tataattaat atcaattggg ttagcaaaac aaatctagtc taggtgtgtt ttgcgaatgc
     3481 ggccgccacc gcggtggagc tcgaattccg gtccgggtca cccagcttga gtattctata
     3541 gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3601 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3661 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3721 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3781 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4381 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4621 agggattttg gtcatggagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     4681 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     4741 gtgttatgag ccatattcaa cgggaaacgt cttgctcgag gccgcgatta aattccaaca
     4801 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     4861 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     4921 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     4981 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     5041 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     5101 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     5161 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     5221 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     5281 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     5341 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     5401 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     5461 tttctccttc attacagaaa cggctttttc agaaatatgg tattgataat cctgatatga
     5521 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     5581 tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt gaataaatcg
     5641 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     5701 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     5761 tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg agtcagcaac
     5821 accttcttca cgagccatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5881 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5941 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6001 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6061 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
     6121 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     6181 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     6241 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     6301 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     6361 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6421 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6481 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6541 cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
     6601 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     6661 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     6721 cgacggccag tgaattgtaa tacgactcac tatagg
//
