LOCUS       sequence-227024-        6070 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 6070)
  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 6070)
  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: pSW179 - pML1
FEATURES             Location/Qualifiers
     source          1..6070
                     /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(5656..5675)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5657..5675)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5685..5707)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5901..5920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6020..6042
                     /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 tagcttatca aagaaaaaac aagaacaaaa
     2281 cgatgcatag tttctaaaat gtgctaaaat tcagaaactg aaacatgatt cattgtctga
     2341 aactttgttt caaattactg aaaataatca ttcactggac caaaacaaat aaataaaata
     2401 aaatcgaatt tctgaatttg gaaattggtt tttggttttt aattttaaac aaaacaaaaa
     2461 cgaaatttga aggcaataaa tgagttagtt ggtaggcaga agtcactcgt tcccactagc
     2521 tattattatt agaagaaacg tccccacaac tccaaggcgt ttcagttcct ttaatttact
     2581 gaattaccct cctcatatct ataaaaaatc acctcttgta ccaatgcccc atttacacat
     2641 cctgtcgttt atttctagac taagtggact acatgtcggt tatttgattc gcaccatgcg
     2701 tatttggatt atcgctaaca caccccttca aacaatacgc ttaactcgta ttacaaaatt
     2761 tcaagtgatg aattatctat gtataagata tagataggaa caactaagca tcgagaaatt
     2821 tgtatataaa tcaactagac ttatatatat ttcgatacag aatttatacg tattatatca
     2881 aattaattag taattgtttc ctctacgtga gtttaattaa caatgataag ctacattgag
     2941 tgtatcagtt ctaaaacttt atagtatgct acaatcaatt tttctaagta acaacttcaa
     3001 gcaaggaatc acacacacac agtggtacat aataaacttg attttaatat catatgatca
     3061 gcatcattaa cggaataagt taagtaattc gtcatccata ctactaagtc atattaaaat
     3121 cataatcaaa cttaaaagcc gattagaaag agagcaaata tatctaaaaa ttcacgagga
     3181 agacgacaaa tgcaaggaaa cacagctagt attattaaac ttaatagata ttggatgaat
     3241 gactgcataa tatatatcac attaaaagtg gacataaatt tgcatatgtg taatgtacct
     3301 ctccacaatt aatcgcggac catttatttt actattacaa gtcaagtaac tttatattgt
     3361 tgatccataa ttcttttcga acataaaatc atatacttag gccattttca actgtcaaaa
     3421 ctcgaatccg agaaccaaat ttcaccattt tccaaaaatg atgagtgtcg accaaatggg
     3481 gtactactgt ctaatcagga acttgtgaac aaattttcaa ccttttccaa ataagacgag
     3541 tgtcaaccaa ctttttccaa ccaagagata ttgggttgct acacaaatac ttaatagcca
     3601 ttgcatattt atgcatatgc aaatgcaggg tcgtggcgtc agaaagaaac ataggaccct
     3661 caacatattt aatattttgg gagctatatt tgactatttc atattagaaa ataataataa
     3721 aaaagtgttg gttttatatc aaattgtaat ttacgaaaaa cttatgcttt tgcgcaatga
     3781 tttttgtaaa gtatctacta tgtttagtgt ttacattgat tagtaggctg ccgttttttt
     3841 tcttgtgtat tatgtactat atatgaatat gaacatttgt aaaagtgaat cttgtcattt
     3901 tcttgttgaa aacatatata gtatgtgcaa acaaagcata ggttaatcca ataccacaca
     3961 aataacacgt caggtaaatc caataataaa tcgtatgtgc atgtatgtgt attcatgtat
     4021 gttacatgaa tgtctgaatc agtcagtgta cgtatatgat gtaggtgatg taaatcttaa
     4081 tgtatgagct gtttcttgga ccatggtcca caatggatat tgctccccaa ctacattagt
     4141 caatcgactg gccaattttt aattaagata attaatccaa actaccatta aatataactt
     4201 tgaccttttt tctattcatt tttagatatt attggaactt acgtagttta catgcatctc
     4261 atccctttct tttgctcctt gaaagtgggt ccaatcacaa aaaatgatct tatattttgt
     4321 attttgtatt ttaaaaactc ataattatat aggttcaaaa atttaattaa catcagtgta
     4381 tactataatt actactctag ccaacaagat aaattcattt tgacatcagc caaaagataa
     4441 aaatttggtt aaaaactatt ggattagctt ttagtattta atattttatg tactgattaa
     4501 atacgaattt agaaatctag gatataagtg agggtgtata ataagggagg ggtggaccat
     4561 taatagcgat gtgcaattaa aaattatgat taagaatcta ggaaatttgt agattgctta
     4621 gttattttta tggcgatcgt cgtgtcaatg tcatggattt tgaaacttta aattaatctc
     4681 ttaaattagc acctaccttt gaattttata gaatcttttt attttatatg tttaatttta
     4741 tagaatctaa ctagcttatt ttgagattaa attgtttagt tacttttata acagtataaa
     4801 tgtataatga ggacctaaga atgtagtcct gtaatgttct tgctattcta cttaatctca
     4861 tcaccaatca accatcaaaa gaagctagta ctaataaaac ctgcaggtat tcgaataata
     4921 attaagctca aacactatac taatttatgg aggattatat attcaatgaa ttaggaacct
     4981 catgatggac attattgact gatataatgt gtatactaat tgtgagtatt taaaaaccat
     5041 acaaagcatt tatatgtcca catatattgg acacacatgc aatcaatgtt caatatgctc
     5101 cacacacaga aataaaaata ctctttctga tcatatgata catcatacat atactaaaaa
     5161 aatctaaaat gaactataac cacaagcata tataataaca atgaaatggt aatgtttctt
     5221 catttttatt tgttcaaatt cttattcggt tgttttttct taccctacga gaatccgtga
     5281 ggtcaaaggg aaacagtgat tttttttttg tattttgttt tttaaattga tgaactgtaa
     5341 aactctctct ctagaaaaat atataagtag tagtatgaat tttctctcac taaaagcatt
     5401 aatggacctt tcgataatca taaatgcaat gcaccctctc tatgcatttc gcaataactc
     5461 cttttccttc tgccacatcc tcttcctcac ctctttctct tcttcccttt ctcctaagtt
     5521 cctcctccac caaattctcc atttatttcg ttaactatcc tccatttgtt ttcttctgaa
     5581 gagtgatata ttctaccttt ctctggttaa agaaactccc tgaatccacc ggttaaacat
     5641 gagaccgaat tctcgcccta tagtgagtcg tattacctgg ccgtcgtttt acaacgtcgt
     5701 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5761 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5821 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5881 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5941 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6001 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6061 aaacgcgcga
//
