LOCUS       sequence-204860-        6775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6775)
  AUTHORS   Chinthalapudi K, Mandati V, Zheng J, Sharff AJ, Bricogne G, Griffin 
            PR, Kissil J, Izard T
  TITLE     Lipid binding promotes the open conformation and tumor-suppressive 
            activity of neurofibromin 2.
  JOURNAL   Nat Commun. 2018 Apr 6;9(1):1338. doi: 10.1038/s41467-018-03648-4.
  PUBMED    29626191
REFERENCE   2  (bases 1 to 6775)
  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: "Nat 
            Commun."; date: "2018-04-6"; volume: "9(1):1338. doi"; pages: " 
            10.1038/s41467-018-03648-4"
COMMENT     Sequence Label: PGEX-6P1-hNF2-AR mutant
FEATURES             Location/Qualifiers
     source          1..6775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      42..630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     531..550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     784..801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        874..951
                     /gene="lacI"
                     /label=lacI promoter
     CDS             952..2034
                     /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     complement(971..990)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2083..2088,2089..2106,2107..2113)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2121..2137
                     /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)."
     primer_bind     2145..2161
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2145..2161
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2157..2684
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2177..2194)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2177..2193)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2186..2208)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2786..2791,2792..2807,2808..2814)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2822..2838
                     /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)."
     CDS             2861..3514
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3472..3494
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3521..3544
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5428..5450)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5595..5613)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5681..5785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5786..5854,5855..6646)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6004..6023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
       61 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      181 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      661 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      721 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
      781 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
      841 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt
      901 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
      961 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     1021 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     1081 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     1141 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     1201 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     1261 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     1321 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     1381 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     1441 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     1501 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     1561 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     1621 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     1681 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     1741 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     1801 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     1861 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     1921 gtgaaaagaa aaaccacgct agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     1981 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2041 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2101 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2161 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2221 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2281 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
     2341 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
     2401 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     2461 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     2521 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     2581 ttatttttga tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg
     2641 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     2701 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     2761 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     2821 attgtgagcg gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta
     2881 ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa
     2941 atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga
     3001 attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca
     3061 gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa
     3121 agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc
     3181 gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc
     3241 tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca
     3301 tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc
     3361 aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc
     3421 acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca
     3481 agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc tgttccaggg
     3541 gcccctggga tccatggccg gggccatcgc ttcccgcatg agcttcagct ctctcaagag
     3601 gaagcaaccc aagacgttca ccgtgaggat cgtcaccatg gacgccgaga tggagttcaa
     3661 ttgcgagatg aagtggaaag ggaaggacct ctttgatttg gtgtgccgga ctctggggct
     3721 ccgagaaacc tggttctttg gactgcagta cacaatcaag gacacagtgg cctggctcaa
     3781 aatggacaag aaggtactgg atcatgatgt ttcaaaggaa gaaccagtca cctttcactt
     3841 cttggccaaa ttttatcctg agaatgctga agaggagctg gttcaggaga tcacacaaca
     3901 tttattcttc ttacaggtaa agaagcagat tttagatgaa aagatctact gccctcctga
     3961 ggcttctgtg ctcctggctt cttacgccgt ccaggccaag tatggtgact acgaccccag
     4021 tgttcacaag cggggatttt tggcccaaga ggaattgctt ccaaaaaggg taataaatct
     4081 gtatcagatg actccggaaa tgtgggagga gagaattact gtttggtacg cagagcaccg
     4141 aggccgagcc agggatgaag ctgaaatgga atatctgaag atagctcagg acctggagat
     4201 gtacggtgtg aactactttg caatccggaa taaaaagggc acagagctgc tgcttggagt
     4261 ggatgccctg gggcttcaca tttatgaccc tgagaacaga ctgaccccca agatctcctt
     4321 cccgtggaat gaaatccgaa acatctcgta cagtgacaag gagtttacta ttaaaccact
     4381 ggataagaaa attgatgtct tcaagtttaa ctcctcaaag cttcgtgtta ataagctgat
     4441 tctccagcta tgtatcgaga accatgatct atttatgagg agaaggaaag ccgattcttt
     4501 ggaagttcag cagatgaaag cccaggccag ggaggagaag gctagaaagc agatggagcg
     4561 gcagcgcctc gctcgagaga agcagatgag ggaggaggct gaacgcacga gggatgagtt
     4621 ggagaggagg ctgctgcaga tgaaagaaga agcaacaatg gccaacgaag cactgatgcg
     4681 gtctgaggag acagctgacc tgttggctga aaaggcccag atcaccgagg aggaggcaaa
     4741 acttctggcc cagaaggccg cagaggctga gcaggaaatg cagcgcatca aggccacagc
     4801 gattcgcacg gaggaggaga agcgcctgat ggagcagaag gtgctggaag ccgaggtgct
     4861 ggcactgaag atggctgagg agtcagagag gagggccaaa gaggcagatc agctgaagca
     4921 ggacctgcag gaagcacgcg aggcggagcg aagagccaag cagaagctcc tggagattgc
     4981 caccaagccc acgtacccgc ccatgaaccc aattccagca ccgttgcctc ctgacatacc
     5041 aagcttcaac ctcattggtg acagcctgtc tttcgacttc aaagatactg acatgaagcg
     5101 gctttccatg gagatagaga aagaaaaagt ggaatacatg gaaaagagca agcatctgca
     5161 ggagcagctc aatgaactca agacagaaat cgaggccttg aaactgaaag agagggagac
     5221 agctctggat attctgcaca atgagaactc cgacagaggt ggcagcagca agcacaatac
     5281 cattaaaaag ctcaccttgc agagctggaa gtccaaagtg gccttctttg aagagctcta
     5341 gcaggtggaa ttcccgggtc gactcgagcg gccgcatcgt gactgactga cgatctgcct
     5401 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5461 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5521 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtataattc
     5581 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5641 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5701 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5761 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5821 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5881 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5941 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6001 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     6061 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6121 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6181 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6241 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6301 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     6361 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6421 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6481 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6541 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6601 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6661 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6721 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttc
//
