LOCUS       sequence-351368-        6217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of PYL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  AUTHORS   Melcher K, Ng LM, Zhou XE, Soon FF, Xu Y, Suino-Powell KM, Park SY, 
            Weiner JJ, Fujii H, Chinnusamy V, Kovach A, Li J, Wang Y, Li J, 
            Peterson FC, Jensen DR, Yong EL, Volkman BF, Cutler SR, Zhu JK, Xu 
            HE
  TITLE     A gate-latch-lock mechanism for hormone signalling by abscisic acid 
            receptors.
  JOURNAL   Nature. 2009 Dec 3;462(7273):602-8. doi: 10.1038/nature08613.
  PUBMED    19898420
REFERENCE   2  (bases 1 to 6217)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2009-12-3- 3"; volume: "462"; issue: "7273"; pages:
            "602-8"
COMMENT     Sequence Label: pSUMO His6 SUMO PYL1 (36-211)
FEATURES             Location/Qualifiers
     source          1..6217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     921..940
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(960..1037)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1243..1262
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1346..1365
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1346..1364
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1365..1389
                     /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)."
     RBS             1404..1426
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1439..1456
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1460..1750
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             2294..2311
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2364..2382)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2378..2425
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2462..2917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2549..2568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2759..2780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2943..3047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3048..3116,3117..3908)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3266..3285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4079..4667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4568..4587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4821..4838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4853..4992
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4918..4937)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5078..5100
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5094..5285)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6094..6217,1..959))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
       61 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
      121 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
      181 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
      241 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
      301 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
      361 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
      421 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
      481 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
      541 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
      601 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
      661 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
      721 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
      781 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
      841 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
      901 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
      961 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     1021 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     1081 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     1141 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     1201 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     1261 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     1321 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
     1381 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata ccatgggtca
     1441 tcaccatcat catcacgggt cggactcaga agtcaatcaa gaagctaagc cagaggtcaa
     1501 gccagaagtc aagcctgaga ctcacatcaa tttaaaggtg tccgatggat cttcagagat
     1561 cttcttcaag atcaaaaaga ccactccttt aagaaggctg atggaagcgt tcgctaaaag
     1621 acagggtaag gaaatggact ccttaagatt cttgtacgac ggtattagaa ttcaagctga
     1681 tcaggcccct gaagatttgg acatggagga taacgatatt attgaggctc accgcgaaca
     1741 gattggaggt ggatccgaat tcacccaact ctcccaatca atcgccgagt tccacacgta
     1801 ccaactcggt aacggccgtt gctcatctct cctagctcag cgaatccacg cgccgccgga
     1861 aacagtatgg tccgtggtga gacgtttcga taggccacag atttacaaac acttcatcaa
     1921 aagctgtaac gtgagtgaag atttcgagat gcgagtggga tgcacgcgcg acgtgaacgt
     1981 gataagtgga ttaccggcga atacgtctcg agagagatta gatctgttgg acgatgatcg
     2041 gagagtgact gggtttagta taaccggtgg tgaacatagg ctgaggaatt ataaatcggt
     2101 tacgacggtt catagatttg agaaagaaga agaagaagaa aggatctgga ccgttgtttt
     2161 ggaatcttat gttgttgatg taccggaagg taattcggag gaagatacga gattgtttgc
     2221 tgatacggtt attagattga atcttcagaa acttgcttcg atcactgaag ctatgaaccg
     2281 gaactaggtc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     2341 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     2401 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     2461 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2521 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2581 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2641 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2701 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2761 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2821 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2881 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     2941 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3001 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3061 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3121 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3181 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3241 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3301 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3361 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3421 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3481 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3541 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     3601 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3661 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3721 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggttctcgc ggtatcattg
     3781 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3841 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3901 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3961 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4021 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4081 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4141 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4201 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4261 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4321 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4381 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4441 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4501 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4561 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4621 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4681 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4741 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4801 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4861 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     4921 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     4981 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5041 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     5101 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     5161 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     5221 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     5281 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     5341 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     5401 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     5461 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     5521 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     5581 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     5641 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     5701 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     5761 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     5821 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     5881 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     5941 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     6001 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     6061 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6121 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6181 gggcgccagg gtggtttttc ttttcaccag tgagacg
//
