LOCUS       sequence-257163-        4432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SthCascade R promoter targeting crRNA2 expression cassette for Zea 
            mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4432)
  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 4432)
  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: PV565
FEATURES             Location/Qualifiers
     source          1..4432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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)."
     primer_bind     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        655..673
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2234..2250)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2235..2251)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2275..2294)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2276..2294)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2301..2318)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2301..2317)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2310..2332)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2459..2914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2546..2565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2756..2777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2940..3044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3045..3113,3114..3905)
                     /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     complement(3263..3282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4076..4432,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tcgaaattaa
      661 ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct ctagaactag
      721 tcagaacaag tacctcggcg tgatcatcca gtgcctcgtg accgagacca agacctccgt
      781 gtccaggcac atctacttct tctccgctcg cggcaggatc gaccccctcg tgtacctcga
      841 cgagttcctc aggaactcag agcccgtgct caagagggtg aacaggaccg gcaactcctc
      901 ctccaacaag caggagtacc agctcctcaa ggacaacctc gtgaggtcct acaacaaggc
      961 cctcaagaag aacgccccct actccatctt cgccatcaag aacggcccca agtcccacat
     1021 cggtaggcac ctcatgacct ccttcctctc aatgaagggc ctcaccgagc tcaccaacgt
     1081 ggtgggcaac tggtccgaca agagggccct gcaggaattc gcggccgcaa aaaaaaggat
     1141 cacccccgcg tgtgcgggaa aaactggttg agtttagctt ctgtaacact actgctggga
     1201 tcacccccgc gtgtgcggga aaaacaattc ggtgcttgcg gctcggttta taaggtacgc
     1261 tccactcttg tgattagtca aggtgggaca aaacgtttgc gtattaccca gcgctttgct
     1321 ccacctacgg actgctgttt gggcttcgtc gcggtgcttc gtattgggcc gtgggccatg
     1381 tttcttatgg ccctctctgt gacgaagttt ttggcacata cgccgctatg actttctctt
     1441 ttactttttt gcaaatattc tccgtcaacg cggcgctgac actgtgtcct actgtacata
     1501 gaaggaaaaa aggtatataa aaaaataatt tctaaacacg acttactgct agtagaaatt
     1561 cgaatagcat tttagctttt acaactgggc ctggaaattc tttggacacc actattttac
     1621 gacttatcat catcgtgtgt tagagcacaa ggaactatac aagcgaggtt gcttatttaa
     1681 gcgggatcat tatgtcaagg attcgatggg gctaacaaaa acagctggaa acacagccag
     1741 atctttgatg ccccctcgct caaaacttgc attccttgta catgcgctaa caaacagaag
     1801 aaatcccttt ttacaataca attttacttt cctattcacc ctgccatata tcatacaaag
     1861 ttggccagga catgcgccca tggccaccat ctctaaccct aatggtgcca tcaaactggc
     1921 accatgatgc cggttatctc actaccttgc tgcaatactg acactctgac agcatgccaa
     1981 tttagctcag cgattactgg tatccttcag ggcctgttcg gttgtaccgg aataagccag
     2041 gattcattcc agctcatcaa aatctatata aattagagaa gtaatccggc taggaattaa
     2101 tccggggctc taatccggaa caaccgaaca gggcctcaag caatcttgat tttctggatc
     2161 atagactgag ggtgcatttt tcagactttg gcattgatta atctaggttc atcattgtac
     2221 tctcaaagct tatcgatacc gtcgacctcg agggggggcc cggtacccaa ttcgccctat
     2281 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2341 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2401 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     2461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2521 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2581 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2641 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2761 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2821 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2881 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2941 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3001 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3061 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3121 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3181 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3241 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3301 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3361 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3421 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3481 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3541 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3601 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3661 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3721 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3781 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3841 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3901 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3961 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4021 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4081 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4141 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4201 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4261 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4321 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4381 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga ac
//
