LOCUS       sequence-350008-        6958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HAB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6958)
  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 6958)
  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: pET24a HAB1 (172-511)
FEATURES             Location/Qualifiers
     source          1..6958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(51..1133)
                     /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"
     primer_bind     1095..1114
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1134..1211)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1417..1436
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1520..1539
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1520..1538
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1539..1563
                     /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             1578..1600
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1620..1637
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1641..2291
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             2298..2315
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3342..3359
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3412..3430)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3426..3473
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3510..3965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3597..3616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3807..3828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4057..4872)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4057..4872)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4780..4799
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4864..4883
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4994..5582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5483..5502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5736..5753
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5768..5907
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5833..5852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5993..6015
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6009..6200)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
       61 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      121 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
      181 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
      241 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
      301 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
      361 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
      421 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
      481 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
      541 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
      601 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
      661 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
      721 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
      781 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
      841 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
      901 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
      961 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     1021 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     1081 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     1141 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     1201 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     1261 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     1321 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     1381 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     1441 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     1501 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
     1561 tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg aaaaaaggtc
     1621 atcatcatca tcatcatggt tcccctatac taggttattg gaaaattaag ggccttgtgc
     1681 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
     1741 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
     1801 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
     1861 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
     1921 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
     1981 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
     2041 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
     2101 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
     2161 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
     2221 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
     2281 atcctccaaa atcggatctg gttccgcgtg gatccaatca tctggtgaaa gggagaagtg
     2341 tttatgaact agattgtata ccgctttggg gcacggtttc cattcaaggg aatagatctg
     2401 agatggagga tgcttttgcc gtgtcacctc attttctgaa actacccatc aaaatgctta
     2461 tgggggacca tgagggtatg agtccaagcc tcacacacct caccggtcat tttttcggtg
     2521 tttatgatgg tcatggaggc cataaggttg ctgactattg ccgagataga ctccattttg
     2581 ctttggctga agaaatagaa cgtataaaag acgaattatg caagaggaat acaggggagg
     2641 gtaggcaggt gcagtgggat aaagtcttca cgagttgttt tctaactgtc gatggtgaga
     2701 ttgaaggaaa aattggtaga gccgttgttg gttcttctga taaggttctt gaggctgttg
     2761 cgtctgagac cgtaggatca actgctgttg ttgccttggt ttgctcatca catatagtag
     2821 tttctaactg cggtgattcg agggcggttt tattccgtgg caaagaagcc atgcccttgt
     2881 cagttgatca caaaccagat agagaggatg aatatgcaag aatagaaaat gctggaggca
     2941 aagttataca atggcaaggc gcacgtgttt ttggtgttct cgccatgtct aggtccatcg
     3001 gtgacagata tctgaagcca tatgtgatcc cagaaccgga agtgacattc atgcctcggt
     3061 caagagaaga cgagtgtctc atactagcca gtgacggtct ttgggatgta atgaacaacc
     3121 aagaagtctg cgaaatagca aggagacgga tattgatgtg gcacaagaag aacggtgcac
     3181 cgcctctagc agagagaggc aaaggaatag atccagcttg ccaagccgca gctgactacc
     3241 tctcaatgct tgctctacaa aaaggaagta aagacaacat ctccatcatt gtgattgact
     3301 tgaaagctca aagaaagttc aagaccagaa cctgactcga gcaccaccac caccaccact
     3361 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     3421 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     3481 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     3541 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3601 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3661 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3721 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3781 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3841 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3901 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3961 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4021 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
     4081 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     4141 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     4201 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     4261 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     4321 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     4381 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     4441 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     4501 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     4561 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     4621 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     4681 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     4741 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     4801 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     4861 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     4921 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4981 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5041 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5101 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5161 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5221 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5281 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5341 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5401 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5461 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5521 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5581 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5641 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5701 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5761 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     5821 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     5881 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     5941 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     6001 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     6061 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     6121 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     6181 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     6241 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     6301 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     6361 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     6421 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     6481 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     6541 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     6601 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     6661 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     6721 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     6781 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     6841 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     6901 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtc
//
