LOCUS       sequence-494295-        6288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A GreenGate entry vector containing coding region of AtREF6 without 
            zinc finger domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6288)
  AUTHORS   Lu J, Jiang D
  TITLE     GreenGate vectors for CRISPR/dCas9-based epigenetic modifiers 
            targeting system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6288)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pGGE-AtREF6-Delta-ZnF
FEATURES             Location/Qualifiers
     source          1..6288
                     /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(5874..5893)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5875..5893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5903..5925)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6119..6138)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6238..6260
                     /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 caaatggctg tgagcgaaca aagccaggac
     2281 gttttcccgt ggctgaagag cctgccggtt gctccggaat tccgtccgac cctggctgag
     2341 tttcaggatc cgatcgctta cattctgaag atcgaggaag aggctagccg ttatggtatt
     2401 tgtaaaatcc tgccgccgct gccgccgccg agcaagaaaa ccagcattag caacctgaat
     2461 cgtagcctgg ctgcgcgtgc tgcggctcgt gttcgtgatg gtggttttgg tgcgtgtgac
     2521 tatgatggtg gtccgacctt cgctacccgt cagcaacaga tcggtttttg cccgcgtaaa
     2581 caacgtccgg tgcagcgtcc ggtttggcag agcggtgaag agtacagctt cggtgaattc
     2641 gagttcaagg ctaagaactt cgaaaagaat tacctgaaga aatgcggtaa gaaaagccaa
     2701 ctgagcgcgc tggaaattga aaccctgtac tggcgtgcta ccgtggacaa accgttcagc
     2761 gttgagtatg cgaacgatat gccgggtagc gcttttatcc cgctgagcct ggctgctgct
     2821 cgtcgtcgtg aaagcggtgg cgagggtggt accgttggtg aaaccgcgtg gaatatgcgt
     2881 gcgatgagcc gtgctgaggg tagcctgctg aagttcatga aagaagagat tccgggtgtt
     2941 accagcccga tggtgtacgt tgcgatgatg ttcagctggt ttgcttggca tgtggaagac
     3001 catgatctgc acagcctgaa ctacctgcac atgggtgcgg gcaagacctg gtatggtgtt
     3061 ccgaaagatg ctgcgctggc ttttgaagag gttgtgcgtg tgcatggtta cggtgaagag
     3121 ctgaacccgc tggttacctt cagcaccctg ggtgaaaaga ccaccgtgat gagcccggag
     3181 gtgtttgtta aagctggtat tccgtgctgt cgtctggttc agaatccggg tgaattcgtt
     3241 gtgacctttc cgggtgcgta tcatagcggt ttcagccacg gtttcaactt tggtgaagcg
     3301 agcaatatcg ctaccccgga gtggctgcgt atggcgaagg acgctgcgat tcgtcgtgct
     3361 gcgatcaatt acccgccgat ggttagccac ctgcaactgc tgtatgattt cgtgctggct
     3421 ctgggtagcc gtgttccgac cagcatcaac ccgaagccgc gtagcagccg tctgaaggat
     3481 aaagcgcgta gcgaaggcga gcgtctgacc aagaaactgt tcgttcagaa catcatccat
     3541 aacaacgaac tgctgagcag cctgggtaaa ggtagcccgg ttgctctgct gccgcaaagc
     3601 agcagcgaca ttagcgtgtg tagcgatctg cgtattggta gccacctgat caccaaccaa
     3661 gaaaatccga tccagctgaa gtgcgaggac ctgagcagcg atagcgttgt ggttgacctg
     3721 agcaacggtc tgaaagatac cgtgagcgtt aaggaaaaat ttaccagcct gtgtgagcgt
     3781 agccgtaatc atctggcgag caccgaaaag gacacccaag aaaccctgag cgatgcggaa
     3841 cgtcgtaaaa acgacgctgc ggttgctctg agcgatcagc gtctgttcag ctgcgtgacc
     3901 tgtggtgttc tgagctttga ctgtgtggcg attgttcaac cgaaagaggc tgcggctcgt
     3961 tatctgatga gcgctgactg cagcttcttt aacgattgga ccgcggctag cggtagcgcg
     4021 aatctgggtc aagcggctcg tagcctgcat ccgcagagca aggagaaaca cgatgttaac
     4081 tacttctata atgtgccggt tcaaaccatg gaccatagcg tgaagaccgg tgatcagaaa
     4141 accagcacca ccagcccgac catcgctcac aaggacaacg atgttctggg catgctggcg
     4201 agcgcttacg gtgacagcag cgatagcgaa gaggaagatc aaaaaggtct ggtgaccccg
     4261 agcagcaagg gtgaaaccaa aacctatgac caggagggta gcgatggtca cgaggaagct
     4321 cgtgacggtc gtaccagcga ttttaactgt caacgtctga ccagcgaaca gaatggtctg
     4381 agcaagggtg gtaaaagcag cctgctggag attgcgctgc cgttcatccc gcgtagcgat
     4441 gacgatagct gtcgtctgca tgtgttttgc ctggaacacg cggctgaagt tgagcaacag
     4501 ctgcgtccgt tcggtggtat taacctgatg ctgctgtgcc atccggagta cccgcgtatt
     4561 gaagcggagg ctaagatcgt ggcggaggaa ctggttatta accacgaatg gaatgacacc
     4621 gagtttcgta atgttacccg tgaagacgag gaaaccatcc aggcggctct ggataacgtg
     4681 gaggctaagg gtggtaatag cgattggacc gtgaaactgg gtgttaacct gagctacagc
     4741 gcgatcctga gccgtagccc gctgtatagc aagcagatgc cgtacaatag catcatttat
     4801 aaagcgttcg gtcgtagcag cccggtggct agcagcccga gcaagccgaa agttagcggc
     4861 aagcgtagca gccgtcaacg taaatatgtg gttggcaagt ggtgtggtaa agtgtggatg
     4921 agccatcaag ttcacccgtt tctgctggaa caggacctgg agggcgagga aagcgaacgt
     4981 agctgccatc tgcgtgttgc gatggacgag gatgctaccg gcaagcgtag cttcccgaac
     5041 aatgtgagcc gtgatagcac caccatgttt ggtcgtaagt actgccgtaa gcgtaaaatt
     5101 cgtgcgaaag ctgttccgcg taagaaactg accagcttca agcgtgaaga cggtgtgagc
     5161 gacgatacca gcgaggatca cagctacaaa caacagtggc gtgctagcgg taacgaggaa
     5221 gagagctatt tcgaaaccgg taacaccgcg agcggtgaca gcagcaatca aatgagcgat
     5281 ccgcataaag gtatcattcg tcacaagggt tacaaagaat ttgagagcga cgatgaagtt
     5341 agcgatcgta gcctgggtga agagtatacc gtgcgtgctt gtgcggctag cgaaagcagc
     5401 atggagaatg gtagccaaca tagcatgtac gaccacgacg atgacgatga cgatattgat
     5461 cgtcagccgc gtggtatccc gcgtagccaa cagacccgtg tgttccgtaa cccggttagc
     5521 tacgaaagcg aggacaatgg tgtttatcaa cagagcggtc gtatcagcat tagcaaccgt
     5581 caggcgaatc gtatggtggg tgaatatgat agcgctgaga acagcctgga agagcgtggt
     5641 ttttgcagca ccggcaagcg tcaaacccgt agcaccgcga aacgtattgc taagaccaaa
     5701 accgttcaaa gcagccgtga caccaaaggt cgtttcctgc aggaatttgc gagcggtaag
     5761 aaaaacgaag agctggatag ctacatggag ggtccgagca cccgtctgcg tgtgcgtcat
     5821 cagaagccga gccgtggtag cctggaaacc aaaactatga gaccgaattc tcgccctata
     5881 gtgagtcgta ttacctggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     5941 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     6001 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     6061 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     6121 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     6181 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6241 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
