LOCUS       sequence-495594-        5755 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  A GreenGate entry vector containing a guide RNA expression cassette 
            targeting the AtFT gene with 7 different gRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5755)
  AUTHORS   Lu J, Jiang D
  TITLE     GreenGate vectors for CRISPR/dCas9-based epigenetic modifiers 
            targeting system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5755)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pGGA-FT guide RNA1_7
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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        2212..2230
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2212..2229
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_RNA        complement(2287..2362)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(2542..2617)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3019..3094)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3496..3571)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3969..4044)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(4441..4516)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4537..4840)
                     /label=AtU6-1 promoter
                     /note="promoter for the Arabidopsis thaliana U6-1 snRNA 
                     gene (Waibel and Filipowicz, 1990)"
     misc_RNA        complement(4896..4971)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4992..5295)
                     /label=AtU6-1 promoter
                     /note="promoter for the Arabidopsis thaliana U6-1 snRNA 
                     gene (Waibel and Filipowicz, 1990)"
     primer_bind     complement(5341..5360)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5342..5360)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5370..5392)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5586..5605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5705..5727
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtggctct cgaggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacgagc tatttaggtg
     2221 acactataga agtgaagctt ggtctcaacc tacgcgtatc catccactcc aagctcttga
     2281 aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac
     2341 ttgctatttc tagctctaaa accgggaagg ccgagattgt agtgaccaat ggtgctttgt
     2401 agccatatat aaagcagtta tttcttcact cgcttaccga tgtgggacgc aaaattcgta
     2461 agaaaaaaga aatcataagc ttattccttt gctgccgatt ccaatcagcg ttctccatcc
     2521 actccaagct cttgaaaaaa agcaccgact cggtgccact ttttcaagtt gataacggac
     2581 tagccttatt ttaacttgct atttctagct ctaaaactgc tacaactgga acaacctgac
     2641 caatgttgct ccctcagtgt tatatataca acaagatttg tttagatttg agcgatgtgg
     2701 gagaaattag gagcttatat gaaacgcata catgggtact gggcatgggt attagtgaat
     2761 ctatggacga tagcccatta aattacataa aaagcccact tatcttattg ctcgatataa
     2821 gctccatata taaactgaag ttgatcctaa aagttcactt gtttcgtcca acatagccaa
     2881 aacagatctt cttaaaccca gacgccattt gtttgattca gttatccatc acaggctgca
     2941 taagaaaaaa tttaaagtaa atcctttgct gccgattcca ttgcagatag tccatccact
     3001 ccaagctctt gaaaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag
     3061 ccttatttta acttgctatt tctagctcta aaacccgaga atatctccat tggtgaccaa
     3121 tgttgctccc tcagtgttat atatacaaca agatttgttt agatttgagc gatgtgggag
     3181 aaattaggag cttatatgaa acgcatacat gggtactggg catgggtatt agtgaatcta
     3241 tggacgatag cccattaaat tacataaaaa gcccacttat cttattgctc gatataagct
     3301 ccatatataa actgaagttg atcctaaaag ttcacttgtt tcgtccaaca tagccaaaac
     3361 agatcttctt aaacccagac gccatttgtt tgattcagtt atccatcaca ggctgcataa
     3421 gaaaaaattt aaagtaaatc ctttgctgcc gattccacaa gtccacatcc atccactcca
     3481 agctcttgaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct
     3541 tattttaact tgctatttct agctctaaaa cttaagatac tctctgctat acaatctctt
     3601 agtcgactct accaatatat aaacagagct actattttca actgaacgat gtgggatgtt
     3661 tttgacatcg tttacagctt aaacgggctc gtttattaaa agccctcttc tttcgatcca
     3721 tcaacattat tggccttaag taaaacaagc ctctttatga ttgagaaaac gccgtcgttg
     3781 gtgcaccaga gagaagaaac aaaacagcta gcgttagata aactttaaaa ttgtttgcat
     3841 cctcatttgt ctgtcaggct gcagtagttt ggattaagaa cgaaactaac tgtaagagat
     3901 ctctgatatt ttcctttgct gccgattcca tgtagtcctt tccatccact ccaagctctt
     3961 gaaaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta
     4021 acttgctatt tctagctcta aaacctcttt ggccataagt aaccaatctc ttagtcgact
     4081 ctaccaatat ataaacagag ctactatttt caactgaacg atgtgggatg tttttgacat
     4141 cgtttacagc ttaaacgggc tcgtttatta aaagccctct tctttcgatc catcaacatt
     4201 attggcctta agtaaaacaa gcctctttat gattgagaaa acgccgtcgt tggtgcacca
     4261 gagagaagaa acaaaacagc tagcgttaga taaactttaa aattgtttgc atcctcattt
     4321 gtctgtcagg ctgcagtagt ttggattaag aacgaaacta actgtaagag atctctgata
     4381 ttttcctttg ctgccgattc caaagtcttc actccatcca ctccaagctc ttgaaaaaaa
     4441 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
     4501 tttctagctc taaaactggt taaaaatgcc ccacgcaatc actacttcgt ctctaaccat
     4561 atataaactc agctgctttc tttacctaag cgctgtggta cttttgaaga ttgttaatag
     4621 cttaaatggg cctattttag aaaaaggcct atccctcgct tccttctcaa gcccatttat
     4681 attattgaat tttttcccaa ctaaagattt tattttctta tgcaagtctc actcacactc
     4741 acgtaaattt caaatataat cgtggtggaa ctaaaacaat gaccgtctcg tttctacgga
     4801 tcgagattca aaattgttct gccggaattt tgagatttct cctttgctgc cgattccagt
     4861 gtcagggtcc atccactcca agctcttgaa aaaaagcacc gactcggtgc cactttttca
     4921 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa cacccgagtt
     4981 aatgcaaatc caatcactac ttcgtctcta accatatata aactcagctg ctttctttac
     5041 ctaagcgctg tggtactttt gaagattgtt aatagcttaa atgggcctat tttagaaaaa
     5101 ggcctatccc tcgcttcctt ctcaagccca tttatattat tgaatttttt cccaactaaa
     5161 gattttattt tcttatgcaa gtctcactca cactcacgta aatttcaaat ataatcgtgg
     5221 tggaactaaa acaatgaccg tctcgtttct acggatcgag attcaaaatt gttctgccgg
     5281 aattttgaga tttctccttt gctgccgatt ccatactagt aacatgagac cgaattctcg
     5341 ccctatagtg agtcgtatta cctggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5401 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5461 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     5521 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     5581 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5641 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5701 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
