LOCUS       sequence-227243-        4979 bp    DNA     circular SYN 15-APR-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 4979)
  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 4979)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pSW377 - pTMO5
FEATURES             Location/Qualifiers
     source          1..4979
                     /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(4565..4584)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4566..4584)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4594..4616)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4810..4829)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4929..4951
                     /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 tgttgaacgt cgtgtgggct tcactctgca
     2281 ccaccacccc aaaaccctag ttcataccct atcctataat ttatactcaa aattttctca
     2341 tttcttctat tatttatcac cattcatatg gaagtatata tttaatgaag aattttcgga
     2401 tctatagatc attatatgta aaatcagttt ttgataaaac atgtgacagt ttcacaaaaa
     2461 tagttcatct ttcacaaaaa tagtactcga atccggtagc attttagtat cctactattt
     2521 agtttttcca ctccttttca ttatatatat aatgatataa gtctttttac ctctttttca
     2581 taagtctttt taactctttt tttctttaaa aatgattata tgcacatcta cttgataaac
     2641 tggcggatgg taaacttgag ttaggtttga gaattttcac taattatttt aggatttgaa
     2701 tcttctttag tatgagacaa ttaagctaac acactcgaag atatattcag atcataatca
     2761 aaatttatcc gttggatatg tatgaataga ttaaataaaa gagagaatct cttcggtgca
     2821 cttgagacaa atctataaca tagaaaatct tgattagatt aaaaactaga ttaaagaagg
     2881 aaagaaaaac aagaaagtgg gtttggctat acgaaacgag tgggattatt aatgattaaa
     2941 agtaaaagtc tttttggtat ctggtcggtc gacaatccgt gtgtcctgtg gtcagtgttt
     3001 gttttttatt cttttggtta attttcgagt attattatgc tttccactga aaaaacaaaa
     3061 acattaagca gccaaatgaa attcttttaa cgcgttgttt cagaacctga tgaaaaaaca
     3121 ataatcatga cctctttctt cttccccttt aaatgcatgt caaattgtgt aggagtgttt
     3181 gattatataa attaaatcta ctttagtctt ctcttctttg tagaattgta gagtcgtggt
     3241 ataaactcga aaatttctta ttatacttat agaaattagg cagacgaaat gctcttgatt
     3301 tgcttacaat cttagattag gtttaaccca agataaaaaa aaactttgtt ttgatgaaga
     3361 aaaacacctt agttacaatt ttggacatca tcttttattt tcacagttag catctgagac
     3421 ttttttttct tttcttaaat agagttctta gacaatatct tttgggtccc tgatcatgtc
     3481 aaatgccaaa aaaataaata agtatacatg attgtgtcta tttgaccatt ctaacttcta
     3541 aaagtttcaa atgaaataac tttgtcaact attatttttg cctatgaatt aggtgctgac
     3601 cagaatcatc ttgctaaaat catgattttc tactattgca tattattctt gatcgcattt
     3661 cacagatctt aatatatacg aaaataactg tatatattat caactgttac gattaatata
     3721 taaccatgca tatcgaaata ttttgaatta gtattttttt tgctaaataa tattttgata
     3781 tagtggtggc gtggtattcc tcatttacaa atcgcatgaa ctttatccta aatacgatta
     3841 acttaagtag aaaaggcaca aaaatcttct attcattaac atgaaatcaa ttcttaagaa
     3901 cataaataaa taataatatt cataggaagc caaatccatc agagaaatcc acaagatgat
     3961 gacagctccc catttagaag tcgtaaaaga aaagcaaaga agatgtgcag cacttgcagt
     4021 ttcatttgtt ttgttagata taccagaaaa aaaaaaagag tggtcccata aaaacctgta
     4081 atatataaca ttttaatcaa agaaaatgat agattttttt attaaacagg aaaaaccact
     4141 aacttttaat taaaaaattt ggaatacagc aatacatcaa aaatccatta acggtacatt
     4201 atagttgaaa ataaattcag gttacataat attttttgcg taatcactat actattctgc
     4261 cattttatat ggaactaagt tcatacgtta tttctcacaa ttctccttga tgtgatcaaa
     4321 gataatggcg gtctgattgt ataacccggt gaaaaaggtt ttttggaaag aattctttaa
     4381 tattaaaata tcaaataaat tttgtttaaa aattaaggac acattatata gcactatata
     4441 tacacatgtg tatgcattat caaatgggat gtttgtgacc ttttgtctct tcttctctaa
     4501 aattctcttt ctcccaaaaa ctaaaaaacc aaaaaaacca aaaaaacatg agaccgaatt
     4561 ctcgccctat agtgagtcgt attacctggc cgtcgtttta caacgtcgtg actgggaaaa
     4621 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4681 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4741 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     4801 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4861 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4921 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
