LOCUS       sequence-494298-        5394 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  A GreenGate entry vector containing coding region of AtCLF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5394)
  AUTHORS   Lu J, Jiang D
  TITLE     GreenGate vectors for CRISPR/dCas9-based epigenetic modifiers 
            targeting system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5394)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pGGE-AtCLF
FEATURES             Location/Qualifiers
     source          1..5394
                     /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"
     primer_bind     complement(4980..4999)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4981..4999)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5009..5031)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5225..5244)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5344..5366
                     /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 ggtctcactg caaatggcgt cagaagcttc gccttcttct
     2281 tcggccacca gatcggagcc acccaaagac tctccggcgg aggagagagg tccagcttct
     2341 aaggaagtat cagaagtaat agaatcgcta aagaagaagc ttgcagctga taggtgtata
     2401 tcaataaaga aaaggattga tgaaaacaag aagaatttgt ttgctattac tcaaagtttt
     2461 atgaggtctt ctatggaacg aggaggtagc tgtaaagatg gcagtgatct tttagttaag
     2521 aggcaaagag attcgccagg tatgaaaagc ggaatcgatg aaagtaataa caacagatat
     2581 gtagaagatg gacctgccag ttcaggaatg gttcaaggat ctagtgtccc tgtcaaaatt
     2641 tccttacgtc ctatcaaaat gcctgatatc aaacgtttgt caccttatac cacatgggtt
     2701 tttctggaca gaaatcaaag aatgactgaa gaccagtctg tagtgggtcg aaggagaatt
     2761 tattatgatc aaactggcgg ggaagcgctt atctgcagtg atagtgaaga ggaggccatt
     2821 gacgacgaag aagaaaaaag agattttttg gagcctgaag attatattat tcgcatgacc
     2881 cttgagcaac taggtctttc agactcagtc ctggcggaac tagcaagttt cttgtctaga
     2941 agtactagtg aaatcaaggc aagacatgga gtgcttatga aggaaaaaga agtatccgag
     3001 agtggcgata atcaagcaga gagctccctt ctcaacaaag atatggaagg agcattagat
     3061 tctttcgata acctgttctg ccgtagatgc cttgtatttg attgccggct tcatgggtgt
     3121 tcacaggatc tcatttttcc ggctgagaaa ccagctccat ggtgtcctcc tgtagatgaa
     3181 aatttaacct gtggtgcaaa ctgctataaa acgcttctca agtctggaag atttccggga
     3241 tatggcacca ttgaaggtaa aactggcact tcatcagatg gtgcaggtac taaaaccaca
     3301 cccacgaagt tctccagcaa acttaatggg agaaaaccaa agaccttccc aagtgaaagt
     3361 gcatcgtcta atgaaaagtg cgcactagaa acaagtgact cagagaatgg actacagcag
     3421 gataccaatt ccgataaagt ttcatcatcg ccaaaggtga aaggtagtgg gagacgagta
     3481 ggtcgtaaga ggaacaaaaa ccgagttgct gagcgagttc ctcgtaagac tcagaagagg
     3541 cagaagaaga cagaagcctc ggatagtgat tccatcgcca gtggaagttg ttcacccagc
     3601 gatgcaaaac ataaagataa tgaagatgct acttcctctt ctcagaagca tgtaaaatct
     3661 gggaactccg ggaagtcaag gaagaatggc actcctgccg aagtctccaa taattctgtg
     3721 aaggatgacg ttcctgtttg ccagtcaaat gaggttgcgt cagagcttga tgcgccgggt
     3781 agtgatgaaa gtctaaggaa agaagagttt atgggtgaaa ctgtatctcg aggaagattg
     3841 gctacaaata agttgtggag gccacttgag aaaagccttt ttgataaagg tgttgagatt
     3901 tttggaatga atagctgctt gattgctaga aatcttttga gtggtttcaa atcatgttgg
     3961 gaggtcttcc aatacatgac gtgctcggaa aataaagctt ccttctttgg aggtgatgga
     4021 ttgaatcctg atggctcttc caagttcgat atcaatggaa atatggttaa taaccaagtg
     4081 aggagaaggt caagatttct acgtaggaga ggcaaagtgc ggcgcttgaa gtatacctgg
     4141 aagtctgctg catatcattc aattaggaaa agaattactg agaagaaaga ccagccctgc
     4201 cgtcagttta atccatgtaa ctgcaaaatt gcttgtggga aggaatgtcc ttgtttgcta
     4261 aacgggactt gctgcgagaa gtactgcggt tgcccaaaga gctgcaagaa taggtttaga
     4321 ggttgtcatt gtgccaaaag tcagtgtcga agccgccagt gtccatgctt tgctgcagat
     4381 cgggaatgtg acccagatgt ttgtagaaac tgctgggtca ttggtgggga tggttcgctt
     4441 ggggtcccaa gccaaagagg cgataattat gagtgcagga atatgaaatt gctcctgaaa
     4501 caacaacaaa gggttttact tggaatatct gatgtttctg gttggggagc tttcttaaag
     4561 aacagtgtaa gtaagcatga ataccttggg gaatacacag gagagctgat ctcacataaa
     4621 gaggcagata aacgcgggaa gatatacgat cgcgagaact gctcttttct cttcaatcta
     4681 aacgatcagt ttgtgctaga tgcttacagg aaaggagata aactgaaatt cgccaaccat
     4741 tctcctgaac ctaactgtta cgcaaaggtc atcatggttg ctggagatca cagggtgggg
     4801 atcttcgcaa aagagaggat actggctgga gaagaactat tttacgatta ccggtatgag
     4861 ccagatcgag ctccagcttg ggccaaaaaa cctgaagctc ctggttctaa gaaagacgaa
     4921 aatgttacac cttctgttgg tagacccaag aagcttgcta ctatgagacc gaattctcgc
     4981 cctatagtga gtcgtattac ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     5041 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     5101 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     5161 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     5221 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5281 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5341 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//
