LOCUS       sequence-227247-        4458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  entry vector for GreenGate cloning method, promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4458)
  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 4458)
  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 
            Physiol. 2018 Jul 19. pii: pp.18.00463. doi: 10.1104/pp.18.00463."
COMMENT     Sequence Label: pSW615 - pATHB-8
FEATURES             Location/Qualifiers
     source          1..4458
                     /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"
     CDS             complement(3374..3385)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4044..4063)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4045..4063)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4073..4095)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4289..4308)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4408..4430
                     /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 tggtcgaaaa atgtataaca atacgcatgc
     2281 aaaattttct ttgcgccaaa gtattaaaat ggcctgcaac tgtacggata aaccaatttt
     2341 caaatgataa tatttgtata taatgtttct gtatacaggg gcggacccac gttagtcttg
     2401 ggtggggcat gtgcccgata ctcaattcta taaatcttat ataagtttgc attatgtaat
     2461 taaaaatgta tttagtttag tggtttttta cttaatggtg ccccacatta acccaagttt
     2521 aaaccttgct gatgtctttt tattgatttt ttataactgt acccatataa gattatttcc
     2581 tggattagcc atctgtatat taataaatcg tttcctgtgg ttgtgcgatg cataaccgtc
     2641 caagaaagtc gtatattttt aatacatgtg ttaaaatagg ttttaagtga gaattttttt
     2701 aacttgtgga gtgtggagtg tagagtgtag atgtacatta cctatatatt tatccaattc
     2761 tgtgtaataa agatgttggg gcggcgcctg ctgccacata cacattggat tagaagggca
     2821 aaagaagttc taataaggag caaactaata gccatgaaaa ttatgaattt accatactga
     2881 aaacaaagga attaaggttt tgaattcaat acttgttttg ttatatatga tgttgatgtg
     2941 acgtatttgt gcgtttctca tagtgacttt gaaaggaagg ctaaacgaat ttgcagtaaa
     3001 agtaacagaa gaaacacttc aaaattgaat taaaatgaag aaaaaaaaga atgagaaaaa
     3061 gtgagaagtg gtggttgtct ggtattaagg gtactcactt ctctcctttt caacacagcc
     3121 cgacacacat gtctcttttt tttttttcat acacacattt ctatttatta gtttcctaaa
     3181 ataaaaatta taaaacaaaa agcacaccca cccataaact aaagcttttc tctctcatcc
     3241 cactacaaat cttatataca agtttcgcaa atcttgactc ttcacgttgt ggatctatct
     3301 agagagataa gtaagagaga ggaagagtaa ggaagaaagt ggttgaactg tagcagagtc
     3361 tgaggtttga atcttgtagt tcccattgaa gcatgcttca ccgttttttc cgaaggaagt
     3421 tactcatcca ctgagagaga aagagggatt tggagataga gaaagaaatt tctcagatgg
     3481 gtttgtgaga attcacagca agagcaagaa actcctatat cttcctttgc ttccagtgac
     3541 cagcgtgatc aaaaacaaat atttttacgg tatctataag aatcctcccc tctttttctt
     3601 tttccttaaa gagaattttt ttgcatcttt cttaggttcc aataaatatg cagaagaaag
     3661 ttaaaatttt tgtaccgtcg ttgaaccttt taaagaccta attaagagac aagatcatac
     3721 taagctcata aatcactttc tatttacaca tatataggtt attaatctta aacaagatct
     3781 atattttatc tttgagattt ttcaagattt catcaagtgt cacgttcatt cattgattac
     3841 atttcaaatt tcactaagaa agttaaaaca cggatctttt tgagaatatc aagaaagttc
     3901 tttgaaatat accaaaagaa atcagtactt tcaagaatac ataacttttt aggttgtgtt
     3961 aaataatatt tgcttttgta ataaaggaac aaaattaaat agagagatcg gagaggatca
     4021 aagaacatga gaccgaattc tcgccctata gtgagtcgta ttacctggcc gtcgttttac
     4081 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4141 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4201 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4261 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4321 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4381 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4441 catcaccgaa acgcgcga
//
