LOCUS       sequence-204861-        7242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7242)
  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 7242)
  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: pcDNA3-Flag-hNF2
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        28..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     656..675
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        656..674
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             686..709
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2588..2606)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2589..2606)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2626..2643)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2632..2856
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2902..3330
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2989..3008)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3199..3220
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3339..3359)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3344..3673
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3524..3659
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3586..3605
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3740..4534
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3794..3813)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4404..4423
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4708..4829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4745..4764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4799..4818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4878..4894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4878..4894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4891..4913)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4902..4918
                     /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        complement(join(4926..4932,4933..4950,4951..4956))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4971..4992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5109..5126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5280..5868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5360..5379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6039..6830,6831..6899))
                     /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     6662..6681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6900..7004)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7079..7098)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      661 cgactcacta tagggagacc ctatggacta caaggacgat gacgataaga agcttggtac
      721 cgagctcgga tccatggccg gggccatcgc ttcccgcatg agcttcagct ctctcaagag
      781 gaagcaaccc aagacgttca ccgtgaggat cgtcaccatg gacgccgaga tggagttcaa
      841 ttgcgagatg aagtggaaag ggaaggacct ctttgatttg gtgtgccgga ctctggggct
      901 ccgagaaacc tggttctttg gactgcagta cacaatcaag gacacagtgg cctggctcaa
      961 aatggacaag aaggtactgg atcatgatgt ttcaaaggaa gaaccagtca cctttcactt
     1021 cttggccaaa ttttatcctg agaatgctga agaggagctg gttcaggaga tcacacaaca
     1081 tttattcttc ttacaggtaa agaagcagat tttagatgaa aagatctact gccctcctga
     1141 ggcttctgtg ctcctggctt cttacgccgt ccaggccaag tatggtgact acgaccccag
     1201 tgttcacaag cggggatttt tggcccaaga ggaattgctt ccaaaaaggg taataaatct
     1261 gtatcagatg actccggaaa tgtgggagga gagaattact gcttggtacg cagagcaccg
     1321 aggccgagcc agggatgaag ctgaaatgga atatctgaag atagctcagg acctggagat
     1381 gtacggtgtg aactactttg caatccggaa taaaaagggc acagagctgc tgcttggagt
     1441 ggatgccctg gggcttcaca tttatgaccc tgagaacaga ctgaccccca agatctcctt
     1501 cccgtggaat gaaatccgaa acatctcgta cagtgacaag gagtttacta ttaaaccact
     1561 ggataagaaa attgatgtct tcaagtttaa ctcctcaaag cttcgtgtta ataagctgat
     1621 tctccagcta tgtatcggga accatgatct atttatgagg agaaggaaag ccgattcttt
     1681 ggaagttcag cagatgaaag cccaggccag ggaggagaag gctagaaagc agatggagcg
     1741 gcagcgcctc gctcgagaga agcagatgag ggaggaggct gaacgcacga gggatgagtt
     1801 ggagaggagg ctgctgcaga tgaaagaaga agcaacaatg gccaacgaag cactgatgcg
     1861 gtctgaggag acagctgacc tgttggctga aaaggcccag atcaccgagg aggaggcaaa
     1921 acttctggcc cagaaggccg cagaggctga gcaggaaatg cagcgcatca aggccacagc
     1981 gattcgcacg gaggaggaga agcgcctgat ggagcagaag gtgctggaag ccgaggtgct
     2041 ggcactgaag atggctgagg agtcagagag gagggccaaa gaggcagatc agctgaagca
     2101 ggacctgcag gaagcacgcg aggcggagcg aagagccaag cagaagctcc tggagattgc
     2161 caccaagccc acgtacccgc ccatgaaccc aattccagca ccgttgcctc ctgacatacc
     2221 aagcttcaac ctcattggtg acagcctgtc tttcgacttc aaagatactg acatgaagcg
     2281 gctttccatg gagatagaga aagaaaaagt ggaatacatg gaaaagagca agcatctgca
     2341 ggagcagctc aatgaactca agacagaaat cgaggccttg aaactgaaag agagggagac
     2401 agctctggat attctgcaca atgagaactc cgacaggggt ggcagcagca agcacaatac
     2461 cattaaaaag ctcaccttgc agagcgccaa gtcccgagtg gccttctttg aagagctcta
     2521 gcaggtggaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg
     2581 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     2641 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2701 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2761 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2821 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2881 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2941 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3001 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3061 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3121 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3181 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3241 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3301 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3361 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3421 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3481 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3541 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3601 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3661 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3721 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3781 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3841 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3901 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3961 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4021 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4081 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4141 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4201 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4261 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4321 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4381 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4441 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4501 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4561 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4621 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4681 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4741 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4801 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4861 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4921 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4981 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5041 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5101 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5161 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5221 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5281 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5341 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5401 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5461 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5521 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5581 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5641 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5701 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5761 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5821 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5881 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5941 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6001 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6061 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6121 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6181 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6241 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6301 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6361 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6421 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6481 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6541 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6601 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6661 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6721 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6781 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6841 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6901 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6961 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7021 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     7081 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7141 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     7201 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tc
//
