LOCUS       sequence-242766-        5460 bp    DNA     circular SYN 14-APR-2025
DEFINITION  intermediate vector for GreenGate cloning method, contains modules 
            from A-F.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5460)
  AUTHORS   Schurholz AK, Lopez-Salmeron V, Li Z, Forner J, Wenzl C, Gaillochet 
            C, Augustin S, Vilches-Barro A, Fuchs M, Gebert M, Lohmann JU, Greb 
            T, Wolf S
  TITLE     A Comprehensive Toolkit for Inducible, Cell Type-Specific Gene 
            Expression in Arabidopsis.
  JOURNAL   Plant Physiol. 2018 Jul 19. pii: pp.18.00463. doi: 
            10.1104/pp.18.00463.
  PUBMED    30026289
REFERENCE   2  (bases 1 to 5460)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol. 2018 Jul 19. pii: pp.18.00463. doi: 10.1104/pp.18.00463."
COMMENT     Sequence Label: pSW609 - N001
FEATURES             Location/Qualifiers
     source          1..5460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    443..462
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    495..514
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    547..566
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    599..618
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        642..688
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             join(773..775,776..778,779..1489)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(818..839)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1079..1098)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1426..1447
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2153..2533
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     primer_bind     complement(3900..3919)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3911..4720
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3978..3997)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4810..5398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5299..5318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggcccggt
      361 ctcatagggg ctatgatgcc tggtcacaac ctatgcatat gtcgagctca agaaatctag
      421 aaagaagaaa gggaagagaa agaattgtga gcgctcacaa ttgaaagact agaaagaaga
      481 aagggaagag aaagaattgt gagcgctcac aattgaaaga ctagaaagaa gaaagggaag
      541 agaaagaatt gtgagcgctc acaattgaaa gactagaaag aagaaaggga agagaaagaa
      601 ttgtgagcgc tcacaattga aagactagtg gatcgatctt cgcaagaccc ttcctctata
      661 taaggaagtt catttcattt ggagaggacg aacaatgaaa gccttcacac tcgctctctt
      721 cttagctctt tccctctatc tcctgcccaa tccagccact agtggctcaa caatggtgag
      781 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      841 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      901 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      961 caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1021 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1081 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1141 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1201 gtacaactac tttagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1261 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1321 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1381 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1441 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt caggtcatga
     1501 tgagctttag ctgcatctcg tctctgttat gcttaagaag ttcaatgttt cgtttcatgt
     1561 aaaactttgg tggtttgtgt tttggggcct tgtataatcc ctgatgaata agtgttctac
     1621 tatgtttccg ttcctgttat ctctttcttt ctaatgacaa gtcgaacttc ttctttatca
     1681 tcgcttcgtt tttattatct gtgcttcttt tgtttaatac gcctgcaaag tgactcgact
     1741 ctgtttagtg cagttctgcg aaacttgtaa atagtccaat tgttggcctc tagtaataga
     1801 tgtagcgaaa gtgttgagct gttgggttct aaggatggct tgaacatgtt aatcttttag
     1861 gttctgagta tgatgaacat tcgttgttgc taagaaatgc ctgtaatgtc ccacaaatgt
     1921 agaaaatggt tcgtaccttt gtccaagcat tgatatgtct gatgagagga aactgcaaga
     1981 tactgagctt ggtttaacga aggagaggca gtttcttcct tccaaagcat ttcatttgac
     2041 aatgccttga tcatcttaag tagagtttct gttgtggaaa gtttgaaact ttgaagaaac
     2101 gactctcaag taaattgatg atcacaagtg aaagtgtatg ttactaggat ccttttcaaa
     2161 tcagtgcgca agacgtgacg taagtatccg agtcagtttt tatttttcta ctaatttggt
     2221 cgtttatttc ggcgtgtagg acatggcaac cgggcctgaa tttcgcgggt attctgtttc
     2281 tattccaact ttttcttgat ccgcagccat taacgacttt tgaatagata cgctgacacg
     2341 ccaagcctcg ctagtcaaaa gtgtaccaaa caacgcttta cagcaagaac ggaatgcgcg
     2401 tgacgctcgc ggtgacgcca tttcgccttt tcagaaatgg ataaatagcc ttgcttccta
     2461 ttatatcttc ccaaattacc aatacattac actagcatct gaatttcata accaatctcg
     2521 atacaccaaa tcgccatggc ttctatgata tcctcttcag ctgtgactac agtcagccgt
     2581 gcttctacgg tgcaatcggc cgcggtggct ccattcggcg gcctcaaatc catgactgga
     2641 ttcccagtta agaaggtcaa cactgacatt acttccatta caagcaatgg tggaagagta
     2701 aagtgcatgg tgacggtgtt cggcattctg aatctcaccg aggactcctt cttcgatgag
     2761 agccggcggc tagaccccgc cggcgctgtc accgcggcga tcgaaatgct gcgagtcgga
     2821 tcagacgtcg tggatgtcgg accggccgcc agccatccgg acgcgaggcc tgtatcgccg
     2881 gccgatgaga tcagacgtat tgcgccgctc ttagacgccc tgtccgatca gatgcaccgt
     2941 gtttcaatcg acagcttcca accggaaacc cagcgctatg cgctcaagcg cggcgtgggc
     3001 tacctgaacg atatccaagg atttcctgac cctgcgctct atcccgatat tgctgaggcg
     3061 gactgcaggc tggtggttat gcactcagcg cagcgggatg gcatcgccac ccgcaccggt
     3121 caccttcgac ccgaagacgc gctcgacgag attgtgcggt tcttcgaggc gcgggtttcc
     3181 gccttgcgac ggagcggggt cgctgccgac cggctcatcc tcgatccggg gatgggattt
     3241 ttcttgagcc ccgcaccgga aacatcgctg cacgtgctgt cgaaccttca aaagctgaag
     3301 tcggcgttgg ggcttccgct acttgtctcg gtgtcgcgga aatccttctt gggcgccacc
     3361 gttggccttc ctgtaaagga tctgggtcca gcgagccttg cggcggaact tcacgcgatc
     3421 ggcaatggcg ctgactacgt ccgcacccac gcgcctggag atctgcgaag cgcaatcacc
     3481 atctcggaaa ccctcgcgaa atttcgcagt cgcgacgcca gagacagagg gttagatcat
     3541 gcctagcttg gctgcaggtc gacctgcagg catgcccgct gaaatcacca gtctctctct
     3601 acaaatctat ctctctctat aataatgtgt gagtagttcc cagataaggg aattagggtt
     3661 cttatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt
     3721 gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag gggtaccgta
     3781 ttgagacccc atccagctgc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     3841 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     3901 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     3961 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     4021 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     4081 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     4141 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     4201 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     4261 tattgttgat gcgctggcag tgtccctgcg ccggttgcat tcgattcctg tttgtaattg
     4321 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     4381 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     4441 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     4501 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     4561 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     4621 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     4681 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     4741 gtaacattat tcagattggg ccccgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
