LOCUS       sequence-258176-        6281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CasB constitutive expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6281)
  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 6281)
  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: PV244
FEATURES             Location/Qualifiers
     source          1..6281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1688..1708
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2097..2115)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2098..2115)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2133..2149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2133..2149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2146..2168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2157..2173
                     /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(2181..2187,2188..2205,2206..2211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2226..2247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2364..2381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2535..3123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2615..2634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3297..3316)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3308..4123
                     /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             3308..4123
                     /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(3381..3400)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4417..4435
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4473..4495)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4595..4614
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4681..5136)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4818..4839)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5030..5049
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5262..5284
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5276..5293
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5277..5293
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5300..5319
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5300..5318
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        join(5353..6281,1..1066)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
ORIGIN
        1 cacacacaca accagatctc ccccaaatcc acccgtcggc acctccgctt caaggtacgc
       61 cgctcgtcct cccccccccc ctctctacct tctctagatc ggcgttccgg tccatgcatg
      121 gttagggccc ggtagttcta cttctgttca tgtttgtgtt agatccgtgt ttgtgttaga
      181 tccgtgctgc tagcgttcgt acacggatgc gacctgtacg tcagacacgt tctgattgct
      241 aacttgccag tgtttctctt tggggaatcc tgggatggct ctagccgttc cgcagacggg
      301 atcgatttca tgattttttt tgtttcgttg catagggttt ggtttgccct tttcctttat
      361 ttcaatatat gccgtgcact tgtttgtcgg gtcatctttt catgcttttt tttgtcttgg
      421 ttgtgatgat gtggtctggt tgggcggtcg ttctagatcg gagtagaatt ctgtttcaaa
      481 ctacctggtg gatttattaa ttttggatct gtatgtgtgt gccatacata ttcatagtta
      541 cgaattgaag atgatggatg gaaatatcga tctaggatag gtatacatgt tgatgcgggt
      601 tttactgatg catatacaga gatgcttttt gttcgcttgg ttgtgatgat gtggtgtggt
      661 tgggcggtcg ttcattcgtt ctagatcgga gtagaatact gtttcaaact acctggtgta
      721 tttattaatt ttggaactgt atgtgtgtgt catacatctt catagttacg agtttaagat
      781 ggatggaaat atcgatctag gataggtata catgttgatg tgggttttac tgatgcatat
      841 acatgatggc atatgcagca tctattcata tgctctaacc ttgagtacct atctattata
      901 ataaacaagt atgttttata attattttga tcttgatata cttggatgat ggcatatgca
      961 gcagctatat gtggattttt ttagccctgc cttcatacgc tatttatttg cttggtactg
     1021 tttcttttgt cgatgctcac cctgttgttt ggtgttactt ctgcaggtcg actctagagg
     1081 atccatgagc cagcatacta aacagaccgt ctggaccgtc accaagaaga tcatcttcaa
     1141 gctcagcaac caaaaggagg aaacctacga caaggccacc ctcgccaagg tcagaaagac
     1201 cctcggcaag cccctcagcg aggccaccga gatctggccc atcctcttcg agaacctccc
     1261 cgaggagttc ctcagcagct accagcaacc aagctacgag gagctcgcca tctacaccgc
     1321 cctccagctc tacgccctcc accagcaggg cgtcagcacc tccgtcatgc tggacgacag
     1381 caagccctat gaaaatatag gcagcgccct cgggaagctg aggaaggagg acgacaccac
     1441 cagcatcgac aggaggttca acaccatgat caccagcagc accttcgagg agctcaccta
     1501 ccacctcagg cacctcatca gcctcctcaa ggtcgagtcc cccgtcatcc aggtcgacta
     1561 cagcggcctc gccaacgacc tctactggtt cctcaataat agccaggaga acgtcaggct
     1621 caactgggcc cgcgcctact acaagcaaat taaaggcgag aagaacaacg acaacagccg
     1681 cgccgacccc aagaagaaga ggaaggtcta gtcttcgtaa gttaacctag acttgtccat
     1741 cttctggatt ggccaactta attaatgtat gaaataaaag gatgcacaca tagtgacatg
     1801 ctaatcacta taatgtgggc atcaaagttg tgtgttatgt gtaattacta gttatctgaa
     1861 taaaagagaa agagatcatc catatttctt atcctaaatg aatgtcacgt gtctttataa
     1921 ttctttgatg aaccagatgc atttcattaa ccaaatccat atacatataa atattaatca
     1981 tatataatta atatcaattg ggttagcaaa acaaatctag tctaggtgtg ttttgcgaat
     2041 gcggccgcca ccgcggtgga gctcgaattc cggtccgggt cacccagctt gagtattcta
     2101 tagtgtcacc taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2161 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2221 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2281 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2341 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2401 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2461 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2701 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2941 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3181 taagggattt tggtcatgga gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     3241 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     3301 gggtgttatg agccatattc aacgggaaac gtcttgctcg aggccgcgat taaattccaa
     3361 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3421 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3481 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3541 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3601 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3661 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3721 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3781 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3841 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3901 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3961 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     4021 gttttctcct tcattacaga aacggctttt tcagaaatat ggtattgata atcctgatat
     4081 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     4141 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcggctttg ttgaataaat
     4201 cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc
     4261 gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac
     4321 cgtggctccc tcactttctg gctggatgat ggggcgattc aggcctggta tgagtcagca
     4381 acaccttctt cacgagccat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4441 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4501 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4561 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4621 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4681 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4741 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4801 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4861 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4921 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4981 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5041 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5101 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa
     5161 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5221 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     5281 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggtt acccggaccg
     5341 aagcttgcat gcctgcagtg cagcgtgacc cggtcgtgcc cctctctaga gataatgagc
     5401 attgcatgtc taagttataa aaaattacca catatttttt ttgtcacact tgtttgaagt
     5461 gcagtttatc tatctttata catatattta aactttactc tacgaataat ataatctata
     5521 gtactacaat aatatcagtg ttttagagaa tcatataaat gaacagttag acatggtcta
     5581 aaggacaatt gagtattttg acaacaggac tctacagttt tatcttttta gtgtgcatgt
     5641 gttctccttt ttttttgcaa atagcttcac ctatataata cttcatccat tttattagta
     5701 catccattta gggtttaggg ttaatggttt ttatagacta atttttttag tacatctatt
     5761 ttattctatt ttagcctcta aattaagaaa actaaaactc tattttagtt tttttattta
     5821 ataatttaga tataaaatag aataaaataa agtgactaaa aattaaacaa atacccttta
     5881 agaaattaaa aaaactaagg aaacattttt cttgtttcga gtagataatg ccagcctgtt
     5941 aaacgccgtc gacgagtcta acggacacca accagcgaac cagcagcgtc gcgtcgggcc
     6001 aagcgaagca gacggcacgg catctctgtc gctgcctctg gacccctctc gagagttccg
     6061 ctccaccgtt ggacttgctc cgctgtcggc atccagaaat tgcgtggcgg agcggcagac
     6121 gtgagccggc acggcaggcg gcctcctcct cctctcacgg caccggcagc tacgggggat
     6181 tcctttccca ccgctccttc gctttccctt cctcgcccgc cgtaataaat agacaccccc
     6241 tccacaccct ctttccccaa cctcgtgttg ttcggagcgc a
//
