LOCUS       Exported                6327 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CasE constitutive expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    PV246
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6327)
  AUTHORS   Joshua K. Young, Stephen L. Gasior, Spencer Jones, Lijuan Wang, 
  TITLE     Repurposing of Type I-E CRISPR-Cascade for gene activation in plants
  JOURNAL   Nature Communications Biology
REFERENCE   2  (bases 1 to 6327)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6327
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1730..1750
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2139..2157)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2140..2157)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2175..2191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2175..2191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2188..2210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2199..2215
                     /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(2223..2253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2268..2289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2406..2423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2577..3165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2657..2676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3339..3358)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3350..4165
                     /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(3423..3442)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4459..4477
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4515..4537)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4637..4656
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4723..5178)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4860..4881)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5072..5091
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5304..5326
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5318..5335
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5319..5335
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5342..5361
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5342..5360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cacacaacca gatctccccc aaatccaccc gtcggcacct ccgcttcaag gtacgccgct
       61 cgtcctcccc cccccccctc tctaccttct ctagatcggc gttccggtcc atgcatggtt
      121 agggcccggt agttctactt ctgttcatgt ttgtgttaga tccgtgtttg tgttagatcc
      181 gtgctgctag cgttcgtaca cggatgcgac ctgtacgtca gacacgttct gattgctaac
      241 ttgccagtgt ttctctttgg ggaatcctgg gatggctcta gccgttccgc agacgggatc
      301 gatttcatga ttttttttgt ttcgttgcat agggtttggt ttgccctttt cctttatttc
      361 aatatatgcc gtgcacttgt ttgtcgggtc atcttttcat gctttttttt gtcttggttg
      421 tgatgatgtg gtctggttgg gcggtcgttc tagatcggag tagaattctg tttcaaacta
      481 cctggtggat ttattaattt tggatctgta tgtgtgtgcc atacatattc atagttacga
      541 attgaagatg atggatggaa atatcgatct aggataggta tacatgttga tgcgggtttt
      601 actgatgcat atacagagat gctttttgtt cgcttggttg tgatgatgtg gtgtggttgg
      661 gcggtcgttc attcgttcta gatcggagta gaatactgtt tcaaactacc tggtgtattt
      721 attaattttg gaactgtatg tgtgtgtcat acatcttcat agttacgagt ttaagatgga
      781 tggaaatatc gatctaggat aggtatacat gttgatgtgg gttttactga tgcatataca
      841 tgatggcata tgcagcatct attcatatgc tctaaccttg agtacctatc tattataata
      901 aacaagtatg ttttataatt attttgatct tgatatactt ggatgatggc atatgcagca
      961 gctatatgtg gattttttta gccctgcctt catacgctat ttatttgctt ggtactgttt
     1021 cttttgtcga tgctcaccct gttgtttggt gttacttctg caggtcgact ctagaggatc
     1081 catgtacatc agcagggtcg agatcgaccg atataatagg cgcaaggtca gggacctcac
     1141 ccacgtcggc gcctaccacg cctgggtcga ggagagcttc cccagcgagc tcgagcaatc
     1201 aatcaggacc aggaagctct ggcgcatcga ccgcatccag ggcaaggact acctcatcat
     1261 cgtcagcaag gagaagccca acctccagaa gctcgaaaaa tatggcgtcg tcggcagcgc
     1321 ccagaccaag gactaccagc acttcctcga cagcatcaag accggcttca ggatgaactt
     1381 caggatcgtc ctcaaccccg tcatcagcat caccagcccc gacaaatcaa aacgcggcgt
     1441 cgtcaagccc cacgtcacca ttaaacacca gatggacttc ctcgtcaagc agagcgaaaa
     1501 aaacggcttc agcctcgtcg gcgagagctt ctccatcgtc gagaggggct acgaggtgtt
     1561 ccagaagccc gagaagccca tcaggctcag caaggtcgtc tacgagggca ccctcaccgt
     1621 cagcgacgac acgctcttca agaagatgct caccgagggc gtcggcaaga agaaagcata
     1681 tggcttcggc ctcatgaccg tcgtccccct cgaggagagc agggccgacc ccaagaagaa
     1741 gaggaaggtc tagtcttcgt aagttaacct agacttgtcc atcttctgga ttggccaact
     1801 taattaatgt atgaaataaa aggatgcaca catagtgaca tgctaatcac tataatgtgg
     1861 gcatcaaagt tgtgtgttat gtgtaattac tagttatctg aataaaagag aaagagatca
     1921 tccatatttc ttatcctaaa tgaatgtcac gtgtctttat aattctttga tgaaccagat
     1981 gcatttcatt aaccaaatcc atatacatat aaatattaat catatataat taatatcaat
     2041 tgggttagca aaacaaatct agtctaggtg tgttttgcga atgcggccgc caccgcggtg
     2101 gagctcgaat tccggtccgg gtcacccagc ttgagtattc tatagtgtca cctaaatagc
     2161 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2221 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2281 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2341 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2401 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2461 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2521 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2581 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2641 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2701 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2761 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2821 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2881 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2941 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3001 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3061 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3241 gagccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3301 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3361 tcaacgggaa acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata
     3421 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     3481 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     3541 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     3601 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     3661 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3721 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3781 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3841 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3901 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3961 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     4021 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     4081 gaaacggctt tttcagaaat atggtattga taatcctgat atgaataaat tgcagtttca
     4141 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     4201 cattacgctg acttgacggg acggcggctt tgttgaataa atcgaacttt tgctgagttg
     4261 aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     4321 aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc
     4381 tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgagcc
     4441 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4501 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4561 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4621 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     4681 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     4741 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4801 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4861 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4921 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4981 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5041 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5101 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5161 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5221 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5281 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     5341 gtaatacgac tcactatagg gcgaattggg ttacccggac cgaagcttgc atgcctgcag
     5401 tgcagcgtga cccggtcgtg cccctctcta gagataatga gcattgcatg tctaagttat
     5461 aaaaaattac cacatatttt ttttgtcaca cttgtttgaa gtgcagttta tctatcttta
     5521 tacatatatt taaactttac tctacgaata atataatcta tagtactaca ataatatcag
     5581 tgttttagag aatcatataa atgaacagtt agacatggtc taaaggacaa ttgagtattt
     5641 tgacaacagg actctacagt tttatctttt tagtgtgcat gtgttctcct ttttttttgc
     5701 aaatagcttc acctatataa tacttcatcc attttattag tacatccatt tagggtttag
     5761 ggttaatggt ttttatagac taattttttt agtacatcta ttttattcta ttttagcctc
     5821 taaattaaga aaactaaaac tctattttag tttttttatt taataattta gatataaaat
     5881 agaataaaat aaagtgacta aaaattaaac aaataccctt taagaaatta aaaaaactaa
     5941 ggaaacattt ttcttgtttc gagtagataa tgccagcctg ttaaacgccg tcgacgagtc
     6001 taacggacac caaccagcga accagcagcg tcgcgtcggg ccaagcgaag cagacggcac
     6061 ggcatctctg tcgctgcctc tggacccctc tcgagagttc cgctccaccg ttggacttgc
     6121 tccgctgtcg gcatccagaa attgcgtggc ggagcggcag acgtgagccg gcacggcagg
     6181 cggcctcctc ctcctctcac ggcaccggca gctacggggg attcctttcc caccgctcct
     6241 tcgctttccc ttcctcgccc gccgtaataa atagacaccc cctccacacc ctctttcccc
     6301 aacctcgtgt tgttcggagc gcacaca
//
