LOCUS       sequence-494255-        6675 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  A GreenGate entry vector containing a ZmUBI promoter and an 
            Insulator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6675)
  AUTHORS   Lu J, Jiang D
  TITLE     GreenGate vectors for CRISPR/dCas9-based epigenetic modifiers 
            targeting system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6675)
  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            "Unpublished"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: pGGC-ZmUBI Promoter-Insulator
FEATURES             Location/Qualifiers
     source          1..6675
                     /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(6261..6280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6262..6280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6290..6312)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6506..6525)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6625..6647
                     /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 tctgcagaag taacaccaaa caacagggtg
     2281 agcatcgaca aaagaaacag taccaagcaa ataaatagcg tatgaaggca gggctaaaaa
     2341 aatccacata tagctgctgc atatgccatc atccaagtat atcaagatca aaataattat
     2401 aaaacatact tgtttattat aatagatagg tactcaaggt tagagcatat gaatagatgc
     2461 tgcatatgcc atcatgtata tgcatcagta aaacccacat caacatgtat acctatccta
     2521 gatcgatatt tccatccatc ttaaactcgt aactatgaag atgtatgaca cacacataca
     2581 gttccaaaat taataaatac accaggtagt ttgaaacagt attctactcc gatctagaac
     2641 gaatgaacga ccgcccaacc acaccacatc atcacaacca agcgaacaaa aagcatctct
     2701 gtatatgcat cagtaaaacc cgcatcaaca tgtataccta tcctagatcg atatttccat
     2761 ccatcatctt caattcgtaa ctatgaatat gtatggcaca cacatacaga tccaaaatta
     2821 ataaatccac caggtagttt gaaacagaat tctactccga tctagaacga ccgcccaacc
     2881 agaccacatc atcacaacca agacaaaaaa aagcatgaaa agatgacccg acaaacaagt
     2941 gcacggcata tattgaaata aaggaaaagg gcaaaccaaa ccctatgcaa cgaaacaaaa
     3001 aaaatcatga aatcgatccc gtctgcggaa cggctagagc catcccagga ttccccaaag
     3061 agaaacactg gcaagttagc aatcagaacg tgtctgacgt acaggtcgca tccgtgtacg
     3121 aacgctagca gcacggatct aacacaaaca cggatctaac acaaacatga acagaagtag
     3181 aactaccggg ccctaaccat ggaccggaac gccgatctag agaaggtaga gagggggggg
     3241 gggaggacga gcggcgtacc ttgaagcgga ggtgccgacg ggtggatttg ggggagatct
     3301 ggttgtgtgt gtgtgcgctc cgaacaacac gaggttgggg aaagagggtg tggagggggt
     3361 gtctatttat tacggcgggc gaggaaggga aagcgaagga gcggtgggaa aggaatcccc
     3421 cgtagctgcc ggtgccgtga gaggaggagg aggccgcctg ccgtgccggc tcacgtctgc
     3481 cgctccgcca cgcaatttct ggatgccgac agcggagcaa gtccaacggt ggagcggaac
     3541 tctcgagagg ggtccagagg cagcgacaga gatgccgtgc cgtctgcttc gcttggcccg
     3601 acgcgacgct gctggttcgc tggttggtgt ccgttagact cgtcgacggc gtttaacagg
     3661 ctggcattat ctactcgaaa caagaaaaat gtttccttag tttttttaat ttcttaaagg
     3721 gtatttgttt aatttttagt cactttattt tattctattt tatatctaaa ttattaaata
     3781 aaaaaactaa aatagagttt tagttttctt aatttagagg ctaaaataga ataaaataga
     3841 tgtactaaaa aaattagtct ataaaaacca ttaaccctaa accctaaatg gatgtactaa
     3901 taaaatggat gaagtattat ataggtgaag ctatttgcaa aaaaaaagga gaacacatgc
     3961 acactaaaaa gataaaactg tagagtcctg ttgtcaaaat actcaattgt cctttagacc
     4021 atgtctaact gttcatttat atgattctct aaaacactga tattattgta gtactataga
     4081 ttatattatt cgtagagtaa agtttaaata tatgtataaa gatagataaa ctgcacttca
     4141 aacaagtgtg acaaaaaaaa tatgtggtaa ttttttataa cttagacatg caatgctcat
     4201 tatctctaga gaggggcacg acttcctaac acctggagaa ccttttatgt acttcacaac
     4261 cctcaatgct gcttccaggt gtgactgttt gggttgctgc atgaattgac tcagcacctg
     4321 cactgcaaag ctgatatctg gccttgtgat tgtcaagtag agaagctttc ccaccagtct
     4381 ctgataggaa cccacatcct ctaactctgc atcatcagtc tttcctaagt gcttgtcata
     4441 ttctacagta gtcagccttt ggttctgttc cattggtgtt gacactggtc tgcagcctcc
     4501 cagaccaaca cctgatatca gttccagtgc atacttcctc tggttcagta ggattccctt
     4561 ttctgatctc agcacctcaa tgcctaagaa gtattttagt tctcccaaat ctttcatctt
     4621 gaaatgctga tgcagggttg cctttgcttc tgaaatcaaa acattgctgc tgcctgttat
     4681 taacagatca tccacataaa tcaggattat gacaaggtca gtcccctctc ttttggtaaa
     4741 caaggagtga tcataagcac tttgcataaa accagcctgc ataaggacag tggtaagctt
     4801 gatgttccac tgccttgatg cttgctttaa accatagagg gattcaacag cctgcacact
     4861 ttggtctgcc cttggctgtg aaaaccctga ggcagagaca tataaacttc ttccatgagg
     4921 tcaccttgta gaaaagcatt gttgacatcc atctggaaaa ggaaccagcc cttggaagca
     4981 gcaacagata tgacagcttt tacagtgacc attttggcca ctggagaaaa agtttcatgg
     5041 tagtcaaggc cttcttgctg agtgtatccc ttggccacta gccttgcctt aaacctgtca
     5101 acttcaccat tagctttgta tttaattttg tacacccatt tggaccctat aggctgttta
     5161 ccagggggta aagggacaat ctcccaggtg ttattatcct caagagcctg tatctcaagg
     5221 gacatggcct ccatccattt ctcatcttga gctgcttctt tgaaggattt aggttcagta
     5281 taagtggaga aagcactcaa ataagcttga tagtgaacag gcaagtgatc ataggaaaca
     5341 tagttggcta tagggtatgg aacatcccta gagcctttgt tcagtgtcac aaagtccttg
     5401 agccagatgg gaggacctgc attgcgttta ggtctagtgt gcaggtttgc tggaactaga
     5461 gagggatcag ctacagcagt atggtgctca aattcagcat ttgctaggtc aggctcagct
     5521 gactcaactg actctgcagg agcatgcagg tggcctgaag gtgcagcatc agctgaagtg
     5581 atatgatttt gggtagaatg agtccccaaa gtgatatcct cattggcctc tacaatgtca
     5641 ggaataagaa ctggagtatc atcatcatca gtatgaaagc tggaaggaaa aatatcatta
     5701 tacactagct gcaaagctgt atcttcagag ctaagtgaac ctgctcgagt gaacatatca
     5761 ggttcatggg agatggactc tttgaaaggg aactgaaact ctctgaaaac tacatccctg
     5821 ctcactatga tcaccttatt atccaagtca tacaacctgt aacccttttg agtctcagaa
     5881 taaccaatga agatggttcg cttggctcta ggtgcaagtt tgtcaccttt gggcagtgtt
     5941 gctgcaaaag caagacaccc aaacactctc aagtgatcaa gcttggcatg tttctgatag
     6001 agtagttcat atggacattt gccttgtaag attggagtag ggagtctatt gatcaggtat
     6061 acaacagtct tgacacagtc tccccaaaac ctggtaggta caccactctg aaacttaagt
     6121 gcccttgcca tctcaaggat gtgtctgtgc tttctctcca caacaccatt ctgttgtggt
     6181 gtgtagggac agctactttg atgaacaatc ccaagagagg ccagcaactc attacaactt
     6241 tcagtgagac cgaattctcg ccctatagtg agtcgtatta cctggccgtc gttttacaac
     6301 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6361 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6421 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     6481 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6541 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6601 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6661 caccgaaacg cgcga
//
