LOCUS       sequence-426176-        6570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning A. thaliana CLV3 promoter as GreenGate module A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6570)
  AUTHORS   Dona M, Bradamante G, Bogojevic Z, Gutzat R, Streubel S, Mosiolek M,
            Dolan L, Mittelsten Scheid O
  TITLE     A versatile CRISPR-based system for lineage tracing in living 
            plants.
  JOURNAL   Plant J. 2023 Jul 5. doi: 10.1111/tpj.16378.
  PUBMED    37403571
REFERENCE   2  (bases 1 to 6570)
  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: "Plant J. 
            2023 Jul 5. doi: 10.1111/tpj.16378."
COMMENT     Sequence Label: pGGA-pCLV3
FEATURES             Location/Qualifiers
     source          1..6570
                     /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(6156..6175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6157..6175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6185..6207)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6401..6420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6520..6542
                     /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 tcgcggtttg tgtaaatggt attattatca
     2281 agatattcta ggcgtggaca taataaatcg aaccgaaaga atcgaattgg aaccaaagta
     2341 aaaagaaccg aactcgaatc cgaaccaaaa tagtaaaata ctcgaatgga tctaaattgt
     2401 agaattataa taattagaat cgaactaaac cgaaccaaaa tatttcaggt atccgaaaaa
     2461 atccaaaata tactcttaaa aataatatta aaatattaaa atatccgaaa aatattcgaa
     2521 aatatccgaa ataaaatatc ctaatggatt ctaaacttta aaaccggaac cgaacatagc
     2581 aaaattgttt tggttcctac aaagctccta aactcgagaa ccgaaagaat cggaaccaaa
     2641 ctagaaccga actggaaccg aatcgaatac ccgaacgccc attcatagct tattaaggcg
     2701 gtaaggctac atgaatgtat gttgtttttt aatttagttt ttggatttga gattttagtt
     2761 ttagatgttg ctattagaac caacccaaaa aaaagtagat tcactaaatt ataagagaac
     2821 taaaaattga agaccttgaa tgtttgtagt ttttaaaatg gttttcgttt ttctgttttt
     2881 ggtctttaga gtaaatttta gtttcttcag aaattgaatc gattgtcacc aaaagattaa
     2941 ttgaatagta tgtgggctca ctatactgac cacgcgaatc tagtgaaggc acactgctac
     3001 tttgaagatg gttatcaaag catcacccta atgcacccta atgctgcaat tccacactag
     3061 tgcgtgctta atccaaccgt agattttcaa agttttaaat tgctttttta gcttaatttt
     3121 tctaatatat gcttgctctt tttaggatgc cacatgaagt tgatctatgg tgagggtggc
     3181 gattaaactt ctcataagga gaagtaagga aaataaactc agctaaaatc ttatactatc
     3241 atacaaattt tagatttcaa ataaaatatt taagtagatt agtttgagaa gaaagtcaaa
     3301 tggcttattt ataagaacaa aaaaaatagc tcgaaaaaat gattaaattt ttgctttgaa
     3361 ttgtgacgct tgctccatca tatgtttgtt agctcattgc atttggctat gcattgataa
     3421 aagttcatga agatgtgaga tactaaattc tcgctcggta atacataagc ttcaactcta
     3481 gctagggttc gtttaccata caattatgat aatccttcat tggtatatct ttttctttta
     3541 tttacttttt tcatccacaa actacattaa cactctatat tctcctccaa atatcaaaac
     3601 tattgatgcc agttttttcg aacacactca tatgttaggt gcttcagttt taaagtttct
     3661 aataacaaaa ctcttagatg tgcatgtaca tgtgggagaa atgaaagtgt gcaagtaatg
     3721 gtgctcatta ttgacgtgaa acttgtcctt aaatttcaaa gaattctacg attttttttt
     3781 gtatttgttg atcaataaga tttttttggt tccaactcta taaaattcat ttactattac
     3841 actcgcataa attacgatga tattatgaaa tgtattaaga ctctaacagt ttaacagtta
     3901 acgacaaaaa tatctactat tgtaaattta aatctttctt tccaactaaa ttgcatattt
     3961 tcggttccgg tataatgaag gcctaaaatc ataagctata ttaatcacat ggaagaatta
     4021 aataaagaaa cctgatcgat atggcagtag aaagaagtca aaagatttgg attttggatt
     4081 gcaacttttt tcttattggt ttatataatt aacaaactag ttgatatata ttatatgctg
     4141 cagaatctta cctccttttt tacatagttt acactttttg tgtgcagctc tcaactcaag
     4201 taggttaatt agacatataa ccaatgaacg taataatatt taaaacttag aattgacctt
     4261 tatgaattca tcaatcagtc tcttgtcgct taacgttaca ctcgtctttt ctggttacta
     4321 ctctcttccc tctcattgca ttttccaggc tgtttcatct ttctttgaca tcactgtatc
     4381 ggtaacgatt cgacgacatc aataaatata ttgttggcca acgaaaaata gacatgagta
     4441 ataatttaat ataatactta ccagtaagtt tacaataatt ttgattgtct tctgatctaa
     4501 taaattgttg gccaaaaaaa gacctaataa attcgtggac ttggagttga tgcagatctt
     4561 tagcagtatg gtggtgttga attcaatctt tttggtccga atgtctttaa aattgttctt
     4621 gatattcata tccataaaat taatcgaatt ccggattatc cataataaaa acaaaactag
     4681 atacattttc taagtacatt aacacataag aatatcattt ttgtactacc aaaaaaaaag
     4741 taaaaactta tgtccatggt attattataa ttggtggtaa atcagtaaat tagttgttag
     4801 caaaaaaata aaatttagtt aaaaagtagt ggcaccttat tggcccaaaa gagtaacgaa
     4861 gcaaaacgga atctgaaaac tgttaaaact taaaagaata gataattgag atataatatc
     4921 gtatgatcgg acggctgtga tctgatgcca tcggatgggc acgtgtcaga agtgtactcc
     4981 aggtatcatt ctctctgcct catgacatca gccgatggta ccctcgatac ggcgccggca
     5041 aggctcatgt aatccattca atttatgttt tttctgctag caaatagatt tgccattgat
     5101 ttgatactct caataatatt atccgattaa ggtatcatcc atattgttca atataattta
     5161 agcatataac tgtttccaga ttaaacaata taatttataa gagcaactgt aatactttac
     5221 tttaaaagtt ttcaaatcag aatctctttt ctttttctac aaatctggaa acaatttaat
     5281 ccatattgtt tttgtttttt ttaccttctc atatttagat gctatttatt attgtgacac
     5341 aagcttagtt taatttcttg ttggttacat ttttttttct aaatacacaa atattatatg
     5401 tttaatatta tttcacacat ttcttagaag aaatctaaat acatgaaatt atatttatta
     5461 aaaagaagtc taaactatat atgtatttaa atggaatttt aaatatgggt agtaatatag
     5521 aaacaccatt gatatattag agtatgtgcc ggtgccgtat acatactgtt gtattatagt
     5581 aacctatata tggtatacct agtcaatgtg tacggtattt ttcatatgta ttagttgtga
     5641 acctccacag cacgttagac ttaggaatta attataactg aaccagaact ttgagtctaa
     5701 ttatacccct gccgttttat acaactttga atattgattc cagtggctaa tttaaataag
     5761 ttaaaggaaa caaaaactgc aagtaagtga aaaatacaca attgtaagta aatgttagat
     5821 atttaattta tttatagcta aatcatgaac aagttcgtat aagatctaga tatatgtacc
     5881 atataccttt tctatgccca ctatatatac ttactacaca taatatataa accaaccaaa
     5941 atatttgaat agttaatagc tatgatacac gtttaggaca agtaaaatta aaaaatagga
     6001 gatcccattt ctcgcccttg taggcttacg ctataaaatt ggactgtccc cttctcattt
     6061 cataaccaaa gtaaagaaca gtttctatat ttctctctgt atctctctca ctcagtcact
     6121 ttctctctaa ggatcaacat gagaccgaat tctcgcccta tagtgagtcg tattacctgg
     6181 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     6241 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     6301 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     6361 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     6421 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6481 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6541 ggttttcacc gtcatcaccg aaacgcgcga
//
