LOCUS       sequence-251964-        8063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  missense mutation in children with EOEE. HA tagged in C-terminus, 
            pCIneo vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8063)
  AUTHORS   Yu W, Shin MR, Sesti F
  TITLE     Complexes formed with integrin-alpha5 and KCNB1 potassium channel 
            wild type or epilepsy-susceptibility variants modulate cellular 
            plasticity via Ras and Akt signaling.
  JOURNAL   FASEB J. 2019 Dec;33(12):14680-14689. doi: 10.1096/fj.201901792R. 
            Epub 2019 Nov 2.
  PUBMED    31682765
REFERENCE   2  (bases 1 to 8063)
  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: 
            "10.1096/fj.201901792R"; journalName: "FASEB J"; date: "2019-12"; 
            volume: "33"; issue: "12"; pages: "14680-14689"
COMMENT     Sequence Label: hKCNB1 L211P
FEATURES             Location/Qualifiers
     source          1..8063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     163..183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     209..233
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          373..505
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     488..507
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        550..568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3174..3200
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3250..3371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3261..3280)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3315..3334
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3557..4012
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3644..3663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3854..3875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4129..4486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4129..4320
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4152..4172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4337..4472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4537..5331
                     /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(4591..4610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5201..5220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5395..5443
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5484..5503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5603..5625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5663..5681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5749..5853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5854..5922,5923..6714)
                     /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(6072..6091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6885..7473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7374..7393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7684..8063
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc
      181 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
      241 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa
      301 cagtctcgaa cttaagctgc agtgactctc ttaaggtagc cttgcagaag ttggtcgtga
      361 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      421 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      481 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      541 tagagtactt aatacgactc actataggct agcctcgaga gctgaattcc accatgccgg
      601 cgggcatgac gaagcatggc tcccgctcca ccagctcgct gccgcccgag cccatggaga
      661 tcgtgcgcag caaggcgtgc tctcggcggg tccgcctcaa cgtcgggggg ctggcgcacg
      721 aggtactctg gcgtaccctg gaccgcctgc cccgcacgcg gctgggcaag ctccgcgact
      781 gcaacacgca cgactcgctg ctggaggtgt gcgatgacta cagcctcgac gacaacgagt
      841 acttctttga ccgccacccg ggcgccttca cctccatcct caacttctac cgcactgggc
      901 gactgcacat gatggaggag atgtgcgcgc tcagcttcag ccaagagctc gactactggg
      961 gcatcgacga gatctacccg gagtcctgct gccaggcccg ctaccaccag aagaaagagc
     1021 agatgaacga ggagctcaag cgtgaggccg agaccctacg ggagcgggaa ggcgaggagt
     1081 tcgataacac gtgctgcgca gagaagagga aaaaactctg ggacctactg gagaagccca
     1141 attcctctgt ggctgccaag atccttgcca taatttccat catgttcatc gtcctctcca
     1201 ccattgccct gtccctcaac acgccgcctg agctacagag cctcgatgag ttcggccagt
     1261 ccacagacaa cccccagctg gcccacgtgg aggccgtgtg catcgcatgg ttcaccatgg
     1321 agtacctgct gaggttcctc tcctcgccca agaagtggaa gttcttcaag ggcccactca
     1381 atgccattga cttgttggcc attctgccat actatgtcac cattttcctc accgaatcca
     1441 acaagagcgt gctgcaattc cagaatgtcc gccgcgtggt ccagatcttc cgcatcatgc
     1501 gcattctccg catccttaag cttgcacgcc actccactgg cctccagtct ctgggcttca
     1561 ctttgcggag gagctacaat gagttgggct tgctcatcct cttccttgcc atgggcatta
     1621 tgatcttctc cagccttgtc ttctttgctg agaaggatga ggacgacacc aagttcaaaa
     1681 gcatcccagc ctctttctgg tgggccacca tcaccatgac tactgttggg tatggagaca
     1741 tctaccccaa gactctcctg gggaaaattg ttgggggact ctgctgcatt gcaggagtcc
     1801 tggtgattgc tcttcccatc cccatcatcg tcaataactt ctctgagttc tataaggagc
     1861 agaagagaca ggagaaagca atcaaacggc gagaggctct ggagagagcc aagaggaatg
     1921 gcagcatcgt atccatgaac atgaaggatg cttttgcccg gagcattgag atgatggaca
     1981 ttgtggttga gaaaaatggg gagaatatgg gtaagaaaga caaagtacaa gataaccact
     2041 tgtctcctaa caaatggaaa tggacaaaga ggacactgtc tgaaaccagc tcaagtaagt
     2101 cctttgaaac caaggaacag ggatcccctg aaaaagccag atcgtcttct agtcctcagc
     2161 acctgaacgt tcagcagttg gaagacatgt acaataagat ggccaagacc caatcccaac
     2221 ccatcctcaa taccaaggag tcagcagcac agagcaaacc aaaggaagaa cttgaaatgg
     2281 agagtatccc cagccccgta gcccctctgc ccactcgcac agaaggggtc attgacatgc
     2341 gaagtatgtc aagcattgat agtttcatta gctgtgccac agacttccct gaggccacca
     2401 gattctccca cagccctttg acatcactcc ccagcaagac tgggggcagc acagccccag
     2461 aagtgggctg gcggggagct ctgggtgcca gtggtggtag gtttgtggag gccaacccca
     2521 gccctgatgc cagccagcac tctagtttct tcatcgagag ccccaagagt tccatgaaaa
     2581 ctaacaaccc tttgaagctc cgagcactta aagtcaactt catggagggt gaccccagtc
     2641 cactcctccc cgttctaggg atgtaccatg accctctcag gaaccggggg agtgctgcgg
     2701 ctgctgtcgc tggactggag tgtgccacgc ttttggacaa ggctgtgctg agcccagagt
     2761 cctccatcta caccacagca agtgctaaga cacccccccg gtctcctgag aaacacacag
     2821 caatagcgtt caactttgag gcgggtgtcc accagtacat tgacgcagac acagatgatg
     2881 agggacagct gctctacagt gtggactcca gcccccccaa aagcctccct gggagcacca
     2941 gtccgaagtt cagcacgggg acaagatcgg agaaaaacca ctttgaaagc tcccctttac
     3001 ccacctcccc taagttctta aggcagaact gtatttactc cacagaagca ttgactggaa
     3061 aaggccccag tggtcaggaa aagtgcaaac ttgagaacca catctcccct gacgtccgtg
     3121 tgttgccagg gggaggagcc catggaagca cacgagatca gagcatcgaa ttctacccat
     3181 acgatgttcc agattacgct taggcggccg cttcccttta gtgagggtta atgcttcgag
     3241 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3301 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3361 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     3421 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatccga taaggatcga
     3481 tccgggctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3541 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3601 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3661 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3721 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3781 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3841 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3901 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3961 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     4021 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     4081 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     4141 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4201 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4261 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4321 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4441 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttgattcttc
     4501 tgacacaaca gtctcgaact taaggctaga gccaccatga ttgaacaaga tggattgcac
     4561 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4621 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4681 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4741 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4801 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4861 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4921 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4981 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5041 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5101 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5161 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5221 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5281 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5341 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa
     5401 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg
     5461 atccgcgtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5521 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5581 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5641 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5701 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5761 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5821 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5881 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5941 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6001 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6061 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6121 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6181 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6241 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6301 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6361 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6421 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6481 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6541 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6601 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6661 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6721 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6781 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6841 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6901 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6961 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7021 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7081 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7141 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7201 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7261 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7321 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7381 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7441 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7501 ttcctggcct tttgctggcc ttttgctcac atggctcgac agatcttcaa tattggccat
     7561 tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     7621 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
     7681 gttggcattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     7741 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     7801 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     7861 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     7921 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
     7981 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
     8041 tattagtcat cgctattacc atg
//
