LOCUS       sequence-494302-        6722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A GreenGate entry vector containing a AtUBQ10 promoter and an 
            Insulator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6722)
  AUTHORS   Lu J, Jiang D
  TITLE     GreenGate vectors for CRISPR/dCas9-based epigenetic modifiers 
            targeting system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6722)
  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            "Unpublished"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: pGGC-AtUBQ10 promoter-Insulator
FEATURES             Location/Qualifiers
     source          1..6722
                     /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(6308..6327)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6309..6327)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6337..6359)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6553..6572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6672..6694
                     /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 ggtctcaggc tccggatccc tgttaatcag aaaaactcag
     2281 attaatcgac aaattcgatc gcacaaacta gaaactaaca ccagatctag atagaaatca
     2341 caaatcgaag agtaattatt cgacaaaact caaattattt gaacaaatcg gatgatatct
     2401 atgaaaccct aatcgagaat taagatgata tctaacgatc aaacccagaa aatcgtcttc
     2461 gatctaagat taacagaatc taaaccaaag aacatatacg aaattgggat cgaacgaaaa
     2521 caaaatcgaa gattttgaga gaataaggaa cacagaaatt taccttgatc acggtagaga
     2581 gaattgagag aaagttttta agattttgag aaattgaaat ctgaattgtg aagaagaaga
     2641 gctctttggg tattgtttta tagaagaaga agaagaaaag acgaggacga ctaggtcacg
     2701 agaaagctaa ggcggtgaag caatagctaa taataaaatg acacgtgtat tgagcgttgt
     2761 ttacacgcaa agttgttttt ggctaattgc cttattttta ggttgaggaa aagtatttgt
     2821 gctttgagtt gataaacacg actcgtgtgt gccggctgca accactttga cgccgtttat
     2881 tactgactcg tcgacaacca caatttctaa cggtcgtcat aagatccagc cgttgagatt
     2941 taacgatcgt tacgatttat atttttttag cattatcgtt ttatttttta aatatacggt
     3001 ggagctgaaa attggcaata attgaaccgt gggtcccact gcattgaagc gtatttcgta
     3061 ttttctagaa ttcttcgtgc tttatttctt ttcctttttg tttttttttg ccatttatct
     3121 aatgcaagtg ggcttataaa atcagtgaat ttcttggaaa agtaacttct ttatcgtata
     3181 acatattgtg aaattatcca tttcttttaa ttttttagtg ttattggata tttttgtatg
     3241 attattgatt tgcataggat aatgactttt gtatcaagtt ggtgaacaag tctcgttaaa
     3301 aaaggcaagt ggtttggtga ctcgatttat tcttgttatt taattcatat atcaatggat
     3361 cttatttggg gcctggtcca tatttaacac tcgtgttcag tccaatgacc aataatattt
     3421 tttcattaat aacaatgtaa caagaatgat acacaaaaca ttctttgaat aagttcgcta
     3481 tgaagaaggg aacttatccg gtcctagatc atcagttcat acaaacctcc atagagttca
     3541 acatcttaaa caagaatatc ctgatctgaa gaatgtggag gctttagtcc cttggatact
     3601 tgggaggctg tggaagaaca gaaacgagct ggtgcttaaa gggagggaat ttggaaccaa
     3661 tgaggtatta gtaaggacac aagaagatgc agatgagtgg attagaagga aagaggctca
     3721 gaatgtaagg aaagcaccaa ccacaacacc gtcgggacag acgagaaaca tgggaagctc
     3781 caccacaatc ttgggttaag tgcaactttg atggggcatg gccaacagaa ggattaaaat
     3841 gtggcttagg gtgggtgctt cgcgatcata cagggaaggt gttatggtta ggtgcaagag
     3901 ctgtggtaaa agtaagaagc gtgctggaag tagaagtgga ggctcttaga tgggctgtgc
     3961 tgtcattatc ccgattcaat tataggaaga tcatttttga ggtggattct cagcaacttg
     4021 tgtctttggt tacatgaaag ttatgcttgt caagtctcaa tccaattatc caagacataa
     4081 agtatctact tagcaagttt gaggatttta tgcttgtgca tacaagccga gaaggaaatg
     4141 gagtggcaga tagaatagct aaggaatctc tttcttttga gaattatgat ctaaagttgt
     4201 attctattgt accaatttgg gttaaaagct ctgttgagct agactcatat gcatgcggcc
     4261 gcgttcctaa cacctggaga accttttatg tacttcacaa ccctcaatgc tgcttccagg
     4321 tgtgactgtt tgggttgctg catgaattga ctcagcacct gcactgcaaa gctgatatct
     4381 ggccttgtga ttgtcaagta gagaagcttt cccaccagtc tctgatagga acccacatcc
     4441 tctaactctg catcatcagt ctttcctaag tgcttgtcat attctacagt agtcagcctt
     4501 tggttctgtt ccattggtgt tgacactggt ctgcagcctc ccagaccaac acctgatatc
     4561 agttccagtg catacttcct ctggttcagt aggattccct tttctgatct cagcacctca
     4621 atgcctaaga agtattttag ttctcccaaa tctttcatct tgaaatgctg atgcagggtt
     4681 gcctttgctt ctgaaatcaa aacattgctg ctgcctgtta ttaacagatc atccacataa
     4741 atcaggatta tgacaaggtc agtcccctct cttttggtaa acaaggagtg atcataagca
     4801 ctttgcataa aaccagcctg cataaggaca gtggtaagct tgatgttcca ctgccttgat
     4861 gcttgcttta aaccatagag ggattcaaca gcctgcacac tttggtctgc ccttggctgt
     4921 gaaaaccctg aggcagagac atataaactt cttccatgag gtcaccttgt agaaaagcat
     4981 tgttgacatc catctggaaa aggaaccagc ccttggaagc agcaacagat atgacagctt
     5041 ttacagtgac cattttggcc actggagaaa aagtttcatg gtagtcaagg ccttcttgct
     5101 gagtgtatcc cttggccact agccttgcct taaacctgtc aacttcacca ttagctttgt
     5161 atttaatttt gtacacccat ttggacccta taggctgttt accagggggt aaagggacaa
     5221 tctcccaggt gttattatcc tcaagagcct gtatctcaag ggacatggcc tccatccatt
     5281 tctcatcttg agctgcttct ttgaaggatt taggttcagt ataagtggag aaagcactca
     5341 aataagcttg atagtgaaca ggcaagtgat cataggaaac atagttggct atagggtatg
     5401 gaacatccct agagcctttg ttcagtgtca caaagtcctt gagccagatg ggaggacctg
     5461 cattgcgttt aggtctagtg tgcaggtttg ctggaactag agagggatca gctacagcag
     5521 tatggtgctc aaattcagca tttgctaggt caggctcagc tgactcaact gactctgcag
     5581 gagcatgcag gtggcctgaa ggtgcagcat cagctgaagt gatatgattt tgggtagaat
     5641 gagtccccaa agtgatatcc tcattggcct ctacaatgtc aggaataaga actggagtat
     5701 catcatcatc agtatgaaag ctggaaggaa aaatatcatt atacactagc tgcaaagctg
     5761 tatcttcaga gctaagtgaa cctgctcgag tgaacatatc aggttcatgg gagatggact
     5821 ctttgaaagg gaactgaaac tctctgaaaa ctacatccct gctcactatg atcaccttat
     5881 tatccaagtc atacaacctg taaccctttt gagtctcaga ataaccaatg aagatggttc
     5941 gcttggctct aggtgcaagt ttgtcacctt tgggcagtgt tgctgcaaaa gcaagacacc
     6001 caaacactct caagtgatca agcttggcat gtttctgata gagtagttca tatggacatt
     6061 tgccttgtaa gattggagta gggagtctat tgatcaggta tacaacagtc ttgacacagt
     6121 ctccccaaaa cctggtaggt acaccactct gaaacttaag tgcccttgcc atctcaagga
     6181 tgtgtctgtg ctttctctcc acaacaccat tctgttgtgg tgtgtaggga cagctacttt
     6241 gatgaacaat cccaagagag gccagcaact cattacaact tggtacctca gtgagaccga
     6301 attctcgccc tatagtgagt cgtattacct ggccgtcgtt ttacaacgtc gtgactggga
     6361 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     6421 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     6481 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     6541 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6601 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6661 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6721 ga
//
