LOCUS       sequence-258178-        5448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C1 over-expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  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 5448)
  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: PHP665
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     251..271
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    329..384
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     primer_bind     complement(2344..2360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2344..2360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2357..2379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2368..2384
                     /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(2392..2398,2399..2416,2417..2422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2437..2458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2575..2592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2746..3334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2826..2845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3505..4296,4297..4365))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4128..4147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4366..4470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4538..4556
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4594..4616)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4716..4735
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4929..4951
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4943..4960
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4944..4960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(5411..5448,1..307)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 tcggattcca ttgcccagct atctgtcact tcatcaaaag gacagtagaa aaggaaggtg
       61 gcacctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat gcctctgccg
      121 acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagacgttc
      181 caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta agggatgacg
      241 cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg
      301 agaggacgac ctgcagctta tttttacaac aattaccaac aacaacaaac aacaaacaac
      361 attacaatta ctatttacaa ttacagtcga cggatcaagt gcaaaggtcc gccttgtttc
      421 tcctctgtct cttgatctga ctaatcttgg tttatgattc gttgagtaat tttggggaaa
      481 gcttcgtcca cagttttttt ttcgatgaac agtgccgcag tggcgctgat cttgtatgct
      541 atcctgcaat cgtggtgaac ttatttcttt tatatccttc actcccatga aaaggctagt
      601 aatctttctc gatgtaacat cgtccagcac tgctattacc gtgtggtcca tccgacagtc
      661 tggctgaaca catcatacga tattgagcaa agatcgatct atcttccctg ttctttaatg
      721 aaagacgtca ttttcatcag tatgatctaa gaatgttgca acttgcaagg aggcgtttct
      781 ttctttgaat ttaactaact cgttgagtgg ccctgtttct cggacgtaag gcctttgctg
      841 ctccacacat gtccattcga attttaccgt gtttagcaag ggcgaaaagt ttgcatcttg
      901 atgatttagc ttgactatgc gattgctttc ctggacccgt gcagctgcgg acggatcccc
      961 gggtaccgag ctcgaattcc gcgcgatggg gaggagggcg tgttgcgcga aggaaggcgt
     1021 taagagaggg gcgtggacga gcaaggagga cgatgccttg gccgcctacg tcaaggccca
     1081 tggcgaaggc aaatggaggg aagtgcccca gaaagccggt ttgcgtcggt gcggcaagag
     1141 ctgccggctg cggtggctga actacctccg gcccaacatc aggcgcggca acatctccta
     1201 cgacgaggag gatctcatca tccgcctcca caggctcctc ggcaacaggt ggtcgctgat
     1261 tgcaggcagg ctgcctggcc gaacagacaa tgaaatcaag aactactgga acagcacgct
     1321 gggccggagg gcaggcgccg gcgccggcgc cggcggcagc tgggtcgtcg tcgcgccgga
     1381 caccggctcg cacgccaccc cggccgcgac gtcgggcgcc tgcgagaccg gccagaatag
     1441 cgccgctcat cgcgcggacc ccgactcagc cgggacgacg acgacctcgg cggcggcggt
     1501 gtgggcgccc aaggccgtgc ggtgcacggg cggactcttc ttcttccacc gggacacgac
     1561 gccggcgcac gcgggcgaga cggcgacgcc aatggccggt ggaggtggag gaggaggagg
     1621 agaagcaggg tcgtcggacg actgcagctc ggcggcgtcg gtatcgcttc gcgtcggaag
     1681 ccacgacgag ccgtgcttct ccggcgacgg tgacggcgac tggatggacg acgtgagggc
     1741 cctggcgtcg tttctcgagt ccgacgagga ctggctccgc tgtcagacgg ccgggcagct
     1801 tgcgtagaca acaagtacac gtatagatgt ccaataagca cgaggcccgc gagcccggca
     1861 cgaagcccgc tttttgggcc cggtccgagc ccggcacggc ccggttatat gcagacccgg
     1921 gccggcccgg cacgaataag cgggccgggc tcggacagga aattaggcac ggtgagctag
     1981 aggatccacc atggttaacc tagacttgtc catcttctgg attggccaac ttaattaatg
     2041 tatgaaataa aaggatgcac acatagtgac atgctaatca ctataatgtg ggcatcaaag
     2101 ttgtgtgtta tgtgtaatta ctagttatct gaataaaaga gaaagagatc atccatattt
     2161 cttatcctaa atgaatgtca cgtgtcttta taattctttg atgaaccaga tgcatttcat
     2221 taaccaaatc catatacata taaatattaa tcatatataa ttaatatcaa ttgggttagc
     2281 aaaacaaatc tagtctaggt gtgttttgcg aatgcggccg cgatctgggg aattcgtaat
     2341 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2401 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2461 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2521 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2581 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2641 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4501 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4561 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4621 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4681 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4741 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4801 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4861 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     4921 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctca gatctgagct
     4981 tctagagatc cgtcaacatg gtggagcacg acactctcgt ctactccaag aatatcaaag
     5041 atacagtctc agaagaccaa agggctattg agacttttca acaaagggta atatcgggaa
     5101 acctcctcgg attccattgc ccagctatct gtcacttcat caaaaggaca gtagaaaagg
     5161 aaggtggcac ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt caagatgcct
     5221 ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag
     5281 acgttccaac cacgtcttca aagcaagtgg attgatgtga tmtctmsasw kmcgtmarsr
     5341 tggtsgmgca cgacactctc gtctactcca agaatatcaa agatacagtc tcagaagacc
     5401 aaagggctat tgagactttt caacaaaggg taatatcggg aaacctcc
//
