LOCUS       sequence-419134-        8205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human MSN in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8205)
  AUTHORS   Du Y, Bradshaw WJ, Leisner TM, Annor-Gyamfi JK, Qian K, Bashore FM, 
            Sikdar A, Nwogbo FO, Ivanov AA, Frye SV, Gileadi O, Brennan PE, 
            Levey AI, Axtman AD, Pearce KH, Fu H, Katis VL
  TITLE     Discovery of FERM domain protein-protein interaction inhibitors for 
            MSN and CD44 as a potential therapeutic approach for Alzheimer's 
            disease.
  JOURNAL   J Biol Chem. 2023 Oct 20:105382. doi: 10.1016/j.jbc.2023.105382.
  PUBMED    37866628
REFERENCE   2  (bases 1 to 8205)
  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: "J Biol 
            Chem. 2023 Oct 20:105382. doi: 10.1016/j.jbc.2023.105382."
COMMENT     Sequence Label: pDEST27-GST-hMSN
FEATURES             Location/Qualifiers
     source          1..8205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     473..493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     519..543
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             632..1285
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1243..1265
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    1313..1337
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3078..3102)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3134..3153)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3135..3153)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3164..3181)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3164..3180)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3173..3195)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    3585..3719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3622..3641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3676..3695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3845..4300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3932..3951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4142..4163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4417..4774
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4417..4608
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4440..4460)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4625..4760
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4825..5619
                     /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(4879..4898)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5489..5508
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5683..5731
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5772..5791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5891..5913
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6037..6141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6142..6210,6211..7002)
                     /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(6360..6379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7173..7761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7662..7681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7915..7932
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(8006..8110)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      601 agcctccgga ctctagccta ggccgcggac catggcccct atactaggtt attggaaaat
      661 taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga
      721 gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt
      781 ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc
      841 catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc
      901 agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc
      961 atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct
     1021 gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca
     1081 tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct
     1141 ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga
     1201 taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt
     1261 tggtggtggc gaccatcctc caaaatcgga tctggttccg cgttctagat caacaagttt
     1321 gtacaaaaaa gcaggctcca ccatgcccaa aacgatcagt gtgcgtgtga ccaccatgga
     1381 tgcagagctg gagtttgcca tccagcccaa caccaccggg aagcagctat ttgaccaggt
     1441 ggtgaaaact attggcttga gggaagtttg gttctttggt ctgcagtacc aggacactaa
     1501 aggtttctcc acctggctga aactcaataa gaaggtgact gcccaggatg tgcggaagga
     1561 aagccccctg ctctttaagt tccgtgccaa gttctaccct gaggatgtgt ccgaggaatt
     1621 gattcaggac atcactcagc gcctgttctt tctgcaagtg aaagagggca ttctcaatga
     1681 tgatatttac tgcccgcctg agaccgctgt gctgctggcc tcgtatgctg tccagtctaa
     1741 gtatggcgac ttcaataagg aagtgcataa gtctggctac ctggccggag acaagttgct
     1801 cccgcagaga gtcctggaac agcacaaact caacaaggac cagtgggagg agcggatcca
     1861 ggtgtggcat gaggaacacc gtggcatgct cagggaggat gctgtcctgg aatatctgaa
     1921 gattgctcaa gatctggaga tgtatggtgt gaactacttc agcatcaaga acaagaaagg
     1981 ctcagagctg tggctggggg tggatgccct gggtctcaac atctatgagc agaatgacag
     2041 actaactccc aagataggct tcccctggag tgaaatcagg aacatctctt tcaatgataa
     2101 gaaatttgtc atcaagccca ttgacaaaaa agccccggac ttcgtcttct atgctccccg
     2161 gctgcggatt aacaagcgga tcttggcctt gtgcatgggg aaccatgaac tatacatgcg
     2221 ccgtcgcaag cctgatacca ttgaggtgca gcagatgaag gcacaggccc gggaggagaa
     2281 gcaccagaag cagatggagc gtgctatgct ggaaaatgag aagaagaagc gtgaaatggc
     2341 agagaaggag aaagagaaga ttgaacggga gaaggaggag ctgatggaga ggctgaagca
     2401 gatcgaggaa cagactaaga aggctcagca agaactggaa gaacagaccc gtagggctct
     2461 ggaacttgag caggaacgga agcgtgccca gagcgaggct gaaaagctgg ccaaggagcg
     2521 tcaagaagct gaagaggcca aggaggcctt gctgcaggcc tcccgggacc agaaaaagac
     2581 tcaggaacag ctggccttgg aaatggcaga gctgacagct cgaatctccc agctggagat
     2641 ggcccgacag aagaaggaga gtgaggctgt ggagtggcag cagaaggccc agatggtaca
     2701 ggaagacttg gagaagaccc gtgctgagct gaagactgcc atgagtacac ctcatgtggc
     2761 agagcctgct gagaatgagc aggatgagca ggatgagaat ggggcagagg ctagtgctga
     2821 cctacgggct gatgctatgg ccaaggaccg cagtgaggag gaacgtacca ctgaggcaga
     2881 gaagaatgag cgtgtgcaga agcacctgaa ggccctcact tcggagctgg ccaatgccag
     2941 agatgagtcc aagaagactg ccaatgacat gatccatgct gagaacatgc gactgggccg
     3001 agacaaatac aagaccctgc gccagatccg gcagggcaac accaagcagc gcattgacga
     3061 atttgagtct atgttggacc cagctttctt gtacaaagtg gttgatcgcg tgcatgcgac
     3121 gtcatagctc tctccctata gtgagtcgta ttataagcta ggcactggcc gtcgttttac
     3181 aacgtcgtga ctgggaaaac tgctagcttg ggatctttgt gaaggaacct tacttctgtg
     3241 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
     3301 ttttaagtgt ataatgtgtt aaactagctg catatgcttg ctgcttgaga gttttgctta
     3361 ctgagtatga tttatgaaaa tattatacac aggagctagt gattctaatt gtttgtgtat
     3421 tttagattca cagtcccaag gctcatttca ggcccctcag tcctcacagt ctgttcatga
     3481 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3541 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3601 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3661 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     3721 atcctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggctggcgt
     3781 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3841 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3901 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3961 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4021 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4081 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4141 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4201 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4261 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     4321 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     4381 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     4441 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4501 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4561 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4621 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4681 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4741 aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt
     4801 ctcgaactta aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg
     4861 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4921 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4981 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5041 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5101 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5161 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5221 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5281 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5341 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5401 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5461 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5521 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5581 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5641 tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt
     5701 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg
     5761 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5821 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5881 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5941 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6001 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6061 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6121 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6181 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6241 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6301 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6361 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6421 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6481 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6541 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6601 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6661 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6721 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6781 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6841 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6901 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6961 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7021 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7081 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7141 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7201 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7261 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7321 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7381 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7441 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7501 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7561 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7621 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7681 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7741 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7801 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7861 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7921 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7981 ccgattcatt aatgcagagc ttgcaattcg cgctttttca atattattga agcatttatc
     8041 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8101 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     8161 tgacattaac ctataaaaat aggcgtatta cgaggccctt tcacg
//
