LOCUS       sequence-254131-        7666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CasA plant transcriptional activator (AT-CBF1) linked constitutive 
            expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7666)
  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 7666)
  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: PV504
FEATURES             Location/Qualifiers
     source          1..7666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     81..103
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     95..112
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     96..112
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     119..138
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        119..137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        172..2167
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             3863..3883
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(4575..4593)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4576..4593)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4611..4627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4611..4627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4624..4646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4635..4651
                     /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    4711..4732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4849..4866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5020..5608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5100..5119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5782..5801)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5793..6608
                     /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             5793..6608
                     /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(5866..5885)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6902..6920
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6958..6980)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7080..7099
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7166..7621)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7303..7324)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7515..7534
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
       61 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
      121 atacgactca ctatagggcg aattgggtta cccggaccga agcttgcatg cctgcagtgc
      181 agcgtgaccc ggtcgtgccc ctctctagag ataatgagca ttgcatgtct aagttataaa
      241 aaattaccac atattttttt tgtcacactt gtttgaagtg cagtttatct atctttatac
      301 atatatttaa actttactct acgaataata taatctatag tactacaata atatcagtgt
      361 tttagagaat catataaatg aacagttaga catggtctaa aggacaattg agtattttga
      421 caacaggact ctacagtttt atctttttag tgtgcatgtg ttctcctttt tttttgcaaa
      481 tagcttcacc tatataatac ttcatccatt ttattagtac atccatttag ggtttagggt
      541 taatggtttt tatagactaa tttttttagt acatctattt tattctattt tagcctctaa
      601 attaagaaaa ctaaaactct attttagttt ttttatttaa taatttagat ataaaataga
      661 ataaaataaa gtgactaaaa attaaacaaa taccctttaa gaaattaaaa aaactaagga
      721 aacatttttc ttgtttcgag tagataatgc cagcctgtta aacgccgtcg acgagtctaa
      781 cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag acggcacggc
      841 atctctgtcg ctgcctctgg acccctctcg agagttccgc tccaccgttg gacttgctcc
      901 gctgtcggca tccagaaatt gcgtggcgga gcggcagacg tgagccggca cggcaggcgg
      961 cctcctcctc ctctcacggc accggcagct acgggggatt cctttcccac cgctccttcg
     1021 ctttcccttc ctcgcccgcc gtaataaata gacaccccct ccacaccctc tttccccaac
     1081 ctcgtgttgt tcggagcgca cacacacaca accagatctc ccccaaatcc acccgtcggc
     1141 acctccgctt caaggtacgc cgctcgtcct cccccccccc cctctctacc ttctctagat
     1201 cggcgttccg gtccatgcat ggttagggcc cggtagttct acttctgttc atgtttgtgt
     1261 tagatccgtg tttgtgttag atccgtgctg ctagcgttcg tacacggatg cgacctgtac
     1321 gtcagacacg ttctgattgc taacttgcca gtgtttctct ttggggaatc ctgggatggc
     1381 tctagccgtt ccgcagacgg gatcgatttc atgatttttt ttgtttcgtt gcatagggtt
     1441 tggtttgccc ttttccttta tttcaatata tgccgtgcac ttgtttgtcg ggtcatcttt
     1501 tcatgctttt ttttgtcttg gttgtgatga tgtggtctgg ttgggcggtc gttctagatc
     1561 ggagtagaat tctgtttcaa actacctggt ggatttatta attttggatc tgtatgtgtg
     1621 tgccatacat attcatagtt acgaattgaa gatgatggat ggaaatatcg atctaggata
     1681 ggtatacatg ttgatgcggg ttttactgat gcatatacag agatgctttt tgttcgcttg
     1741 gttgtgatga tgtggtgtgg ttgggcggtc gttcattcgt tctagatcgg agtagaatac
     1801 tgtttcaaac tacctggtgt atttattaat tttggaactg tatgtgtgtg tcatacatct
     1861 tcatagttac gagtttaaga tggatggaaa tatcgatcta ggataggtat acatgttgat
     1921 gtgggtttta ctgatgcata tacatgatgg catatgcagc atctattcat atgctctaac
     1981 cttgagtacc tatctattat aataaacaag tatgttttat aattattttg atcttgatat
     2041 acttggatga tggcatatgc agcagctata tgtggatttt tttagccctg ccttcatacg
     2101 ctatttattt gcttggtact gtttcttttg tcgatgctca ccctgttgtt tggtgttact
     2161 tctgcaggtc gactctagag gatccatgag ccgcttcaac ctcctcgacg agccctggat
     2221 cagcgtcatc tacgacgaga agggcagcac caaggacgtc agcctccagg acctcttcac
     2281 caacgcccac cagtacaagg agctcgccgg cgacaccaag acccaggact tcgccgtcct
     2341 cagggtcctc ctcgccgtcc tccacaccgt gttcagccgc ttcgacatcg acggcaacgc
     2401 ctacgagtac ctcaccatcg acgagggctg gaaccagctc gagcccgtcg acgagatgga
     2461 catcgagaac tacgaggagg ccctctatga aacatgggag aagctctgga ccaataaaag
     2521 attccccaac atcgtcaatc aatacctcga gaagtggagg gaccgcttct acctcttcga
     2581 ccagaagtac cccttcttcc aggtcaccaa ggaggacatc gccggcgata aaattagcaa
     2641 ggccaagggc accagcatcc tcggcaaaaa tataaatcgc atcatcagcg agagcggcaa
     2701 caagatcgcc ctcttcagcc ccaaggacga ggaaaataaa aatactctca ccgccgccga
     2761 gctcgccagg tggctcatca ccttccaggg ctacagcggc gtcagcgaca aggtcatctt
     2821 cggcaatgaa aaatataccc ccagcaaggg ctggctcttc gacatcggcg ccatctacat
     2881 taaaggcaac accctcttcg aaaccctcct cctcaactac atcagcccct acaacgagtg
     2941 cggcaatgtg gaaaatatac agaggccctg ctgggagagg aagagcagcg acatcattaa
     3001 aagctacctc gatgaaaaag acatcaccaa catcgccagc ctctacaccg tctggagcaa
     3061 gggcatctac atcgaccccg acttcaacct caataaaccc ttcagcttcg acatcgtcaa
     3121 gctccccgac atcaaccacc gcgacaactt cctcgagccc atgaccctct ggaagtacaa
     3181 cgtcagcggc aaaaataggg acagctacac ccccaagaag cacctcctca accagagcct
     3241 ctggaggagc ttcggcctcc tgagcataaa agacaccgac ctccagcaca ggaagcccgg
     3301 cgtcatcgag tggctcacct acatcgacga tataataggc aacaggctca gcaacatcgt
     3361 cgccatcagc atgcaggacg acggcaaccc ccagagcagg ctgcccaccg acgaggtcat
     3421 cgacagcatc ttcatcaacg acctcgtcct caccgacctc gacgagggcg gctgggtccc
     3481 caggatcaac gaggtcgtcg aggaaaccaa gaagatcatc agcaggacat ataaaacgta
     3541 cattaatgat ataaaggaga tccgcaacat cagcaacggc agctacaccc agcagatcgt
     3601 cgagagcctc tacttcaaga tcgaccagcc cttcaggcag tggctcgcca gcatccagcc
     3661 cgaggacgac aaggacagca agatccagga gtggcgcgtc ctcctcaaga aaatagtcaa
     3721 ggccgaggcc gaggtcatcc tccaccaggg cggcgccagg gactacctcg gcatccagaa
     3781 ggacggccga attaagaaca tcgccaccgc ctacaacagc ttcgacttct ggctccgcca
     3841 gcagttaaaa agcagggccg accccaagaa gaagaggaag gtgggtcggg cgacatgtgc
     3901 caaggacatc cagaaggccg ctgccgaagc ggcgctcgct tttcaggacg aaacctgcga
     3961 caccaccacc accaaccacg gcctcgacat ggaggaaaca atggtggagg ccatctacac
     4021 gcctgaacag agcgaggggg ctttctacat ggacgaggaa accatgttcg gcatgcccac
     4081 actcctcgac aacatggccg agggcatgct cctccccccc ccgagcgtcc agtggaacca
     4141 caactacgac ggcgaaggcg acggcgacgt cagcctctgg tcctactagt cttcgtaagt
     4201 taacctagac ttgtccatct tctggattgg ccaacttaat taatgtatga aataaaagga
     4261 tgcacacata gtgacatgct aatcactata atgtgggcat caaagttgtg tgttatgtgt
     4321 aattactagt tatctgaata aaagagaaag agatcatcca tatttcttat cctaaatgaa
     4381 tgtcacgtgt ctttataatt ctttgatgaa ccagatgcat ttcattaacc aaatccatat
     4441 acatataaat attaatcata tataattaat atcaattggg ttagcaaaac aaatctagtc
     4501 taggtgtgtt ttgcgaatgc ggccgccacc gcggtggagc tcgaattccg gtccgggtca
     4561 cccagcttga gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg
     4621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4681 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4741 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4801 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4861 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4921 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5461 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atggagccac gttgtgtctc
     5701 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     5761 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     5821 gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     5881 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     5941 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     6001 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     6061 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     6121 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     6181 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     6241 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     6301 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     6361 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     6421 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     6481 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaga
     6541 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     6601 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     6661 gggacggcgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     6721 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     6781 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc
     6841 gattcaggcc tggtatgagt cagcaacacc ttcttcacga gccatgacat taacctataa
     6901 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6961 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7021 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7081 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     7141 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
     7201 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     7261 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     7321 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     7381 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     7441 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     7501 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     7561 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7621 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggt
//
