LOCUS       sequence-257165-        4438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 DsRed targeting sgRNA expression cassette for Zea mays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4438)
  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 4438)
  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: PV384
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(15..20,21..38,39..45)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    53..69
                     /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     58..80
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     77..93
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     77..93
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        114..132
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        complement(597..672)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1699..1715)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1700..1716)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1740..1759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1741..1759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1766..1783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1766..1782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1775..1797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1924..2379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2011..2030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2221..2242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2405..2509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2510..2578,2579..3370)
                     /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(2728..2747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3541..4129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4030..4049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4283..4300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4417..4438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
       61 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac
      121 cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggccgctc tagaactagt
      181 cagaacaagt acctcggcgt gatcatccag tgcctcgtga ccgagaccaa gacctccgtg
      241 tccaggcaca tctacttctt ctccgctcgc ggcaggatcg accccctcgt gtacctcgac
      301 gagttcctca ggaactcaga gcccgtgctc aagagggtga acaggaccgg caactcctcc
      361 tccaacaagc aggagtacca gctcctcaag gacaacctcg tgaggtccta caacaaggcc
      421 ctcaagaaga acgcccccta ctccatcttc gccatcaaga acggccccaa gtcccacatc
      481 ggtaggcacc tcatgacctc cttcctctca atgaagggcc tcaccgagct caccaacgtg
      541 gtgggcaact ggtccgacaa gagggccctg caggaattcg cggccgcaaa aaaaaagcac
      601 cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac ttgctatttc
      661 tagctctaaa acgattttct ttctgtcctc aattcggtgc ttgcggctcg gtttataagg
      721 tacgctccac tcttgtgatt agtcaaggtg ggacaaaacg tttgcgtatt acccagcgct
      781 ttgctccacc tacggactgc tgtttgggct tcgtcgcggt gcttcgtatt gggccgtggg
      841 ccatgtttct tatggccctc tctgtgacga agtttttggc acatacgccg ctatgacttt
      901 ctcttttact tttttgcaaa tattctccgt caacgcggcg ctgacactgt gtcctactgt
      961 acatagaagg aaaaaaggta tataaaaaaa taatttctaa acacgactta ctgctagtag
     1021 aaattcgaat agcattttag cttttacaac tgggcctgga aattctttgg acaccactat
     1081 tttacgactt atcatcatcg tgtgttagag cacaaggaac tatacaagcg aggttgctta
     1141 tttaagcggg atcattatgt caaggattcg atggggctaa caaaaacagc tggaaacaca
     1201 gccagatctt tgatgccccc tcgctcaaaa cttgcattcc ttgtacatgc gctaacaaac
     1261 agaagaaatc cctttttaca atacaatttt actttcctat tcaccctgcc atatatcata
     1321 caaagttggc caggacatgc gcccatggcc accatctcta accctaatgg tgccatcaaa
     1381 ctggcaccat gatgccggtt atctcactac cttgctgcaa tactgacact ctgacagcat
     1441 gccaatttag ctcagcgatt actggtatcc ttcagggcct gttcggttgt accggaataa
     1501 gccaggattc attccagctc atcaaaatct atataaatta gagaagtaat ccggctagga
     1561 attaatccgg ggctctaatc cggaacaacc gaacagggcc tcaagcaatc ttgattttct
     1621 ggatcataga ctgagggtgc atttttcaga ctttggcatt gattaatcta ggttcatcat
     1681 tgtactctca aagcttatcg ataccgtcga cctcgagggg gggcccggta cccaattcgc
     1741 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1801 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     1861 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1921 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1981 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2041 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2101 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2161 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2221 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2281 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2341 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     2401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2521 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2581 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2641 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2701 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2761 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2821 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2881 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2941 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3001 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3061 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3121 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3181 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3241 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3301 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3361 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3421 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3481 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3541 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3601 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3661 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3721 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3781 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3841 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3901 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3961 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4021 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4081 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4141 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4201 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4261 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4321 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4381 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcat
//
