LOCUS       sequence-251791-        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 R312H
FEATURES             Location/Qualifiers
     source          1..8063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3554..3580
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3630..3751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3641..3660)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3695..3714
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3937..4392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4024..4043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4234..4255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4509..4866
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4509..4700
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4532..4552)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4717..4852
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4917..5711
                     /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(4971..4990)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5581..5600
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5775..5823
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5864..5883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5983..6005
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6043..6061)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6129..6233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6234..6302,6303..7094)
                     /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(6452..6471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7265..7853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7754..7773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcctcgaga
      961 gctgaattcc accatgccgg cgggcatgac gaagcatggc tcccgctcca ccagctcgct
     1021 gccgcccgag cccatggaga tcgtgcgcag caaggcgtgc tctcggcggg tccgcctcaa
     1081 cgtcgggggg ctggcgcacg aggtactctg gcgtaccctg gaccgcctgc cccgcacgcg
     1141 gctgggcaag ctccgcgact gcaacacgca cgactcgctg ctggaggtgt gcgatgacta
     1201 cagcctcgac gacaacgagt acttctttga ccgccacccg ggcgccttca cctccatcct
     1261 caacttctac cgcactgggc gactgcacat gatggaggag atgtgcgcgc tcagcttcag
     1321 ccaagagctc gactactggg gcatcgacga gatctacccg gagtcctgct gccaggcccg
     1381 ctaccaccag aagaaagagc agatgaacga ggagctcaag cgtgaggccg agaccctacg
     1441 ggagcgggaa ggcgaggagt tcgataacac gtgctgcgca gagaagagga aaaaactctg
     1501 ggacctactg gagaagccca attcctctgt ggctgccaag atccttgcca taatttccat
     1561 catgttcatc gtcctctcca ccattgccct gtccctcaac acgctgcctg agctacagag
     1621 cctcgatgag ttcggccagt ccacagacaa cccccagctg gcccacgtgg aggccgtgtg
     1681 catcgcatgg ttcaccatgg agtacctgct gaggttcctc tcctcgccca agaagtggaa
     1741 gttcttcaag ggcccactca atgccattga cttgttggcc attctgccat actatgtcac
     1801 cattttcctc accgaatcca acaagagcgt gctgcaattc cagaatgtcc gccgcgtggt
     1861 ccagatcttc cgcatcatgc gaattctccg catccttaag cttgcacacc actccactgg
     1921 cctccagtct ctgggcttca ctttgcggag gagctacaat gagttgggct tgctcatcct
     1981 cttccttgcc atgggcatta tgatcttctc cagccttgtc ttctttgctg agaaggatga
     2041 ggacgacacc aagttcaaaa gcatcccagc ctctttctgg tgggccacca tcaccatgac
     2101 tactgttggg tatggagaca tctaccccaa gactctcctg gggaaaattg ttgggggact
     2161 ctgctgcatt gcaggagtcc tggtgattgc tcttcccatc cccatcatcg tcaataactt
     2221 ctctgagttc tataaggagc agaagagaca ggagaaagca atcaaacggc gagaggctct
     2281 ggagagagcc aagaggaatg gcagcatcgt atccatgaac atgaaggatg cttttgcccg
     2341 gagcattgag atgatggaca ttgtggttga gaaaaatggg gagaatatgg gtaagaaaga
     2401 caaagtacaa gataaccact tgtctcctaa caaatggaaa tggacaaaga ggacactgtc
     2461 tgaaaccagc tcaagtaagt cctttgaaac caaggaacag ggatcccctg aaaaagccag
     2521 atcgtcttct agtcctcagc acctgaacgt tcagcagttg gaagacatgt acaataagat
     2581 ggccaagacc caatcccaac ccatcctcaa taccaaggag tcagcagcac agagcaaacc
     2641 aaaggaagaa cttgaaatgg agagtatccc cagccccgta gcccctctgc ccactcgcac
     2701 agaaggggtc attgacatgc gaagtatgtc aagcattgat agtttcatta gctgtgccac
     2761 agacttccct gaggccacca gattctccca cagccctttg acatcactcc ccagcaagac
     2821 tgggggcagc acagccccag aagtgggctg gcggggagct ctgggtgcca gtggtggtag
     2881 gtttgtggag gccaacccca gccctgatgc cagccagcac tctagtttct tcatcgagag
     2941 ccccaagagt tccatgaaaa ctaacaaccc tttgaagctc cgagcactta aagtcaactt
     3001 catggagggt gaccccagtc cactcctccc cgttctaggg atgtaccatg accctctcag
     3061 gaaccggggg agtgctgcgg ctgctgtcgc tggactggag tgtgccacgc ttttggacaa
     3121 ggctgtgctg agcccagagt cctccatcta caccacagca agtgctaaga cacccccccg
     3181 gtctcctgag aaacacacag caatagcgtt caactttgag gcgggtgtcc accagtacat
     3241 tgacgcagac acagatgatg agggacagct gctctacagt gtggactcca gcccccccaa
     3301 aagcctccct gggagcacca gtccgaagtt cagcacgggg acaagatcgg agaaaaacca
     3361 ctttgaaagc tcccctttac ccacctcccc taagttctta aggcagaact gtatttactc
     3421 cacagaagca ttgactggaa aaggccccag tggtcaggaa aagtgcaaac ttgagaacca
     3481 catctcccct gacgtccgtg tgttgccagg gggaggagcc catggaagca cacgagatca
     3541 gagcatcgaa ttctacccat acgatgttcc agattacgct taggcggccg cttcccttta
     3601 gtgagggtta atgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3661 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3721 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3781 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3841 taaaatccga taaggatcga tccgggctgg cgtaatagcg aagaggcccg caccgatcgc
     3901 ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
     3961 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4021 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4081 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4141 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4201 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4261 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4321 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4381 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4441 catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     4501 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     4561 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4621 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4681 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4741 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4801 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4861 tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taaggctaga gccaccatga
     4921 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4981 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5041 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5101 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5161 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5221 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5281 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5341 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5401 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5461 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5521 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5581 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5641 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5701 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5761 atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     5821 gtgaatcgat agcgataagg atccgcgtat ggtgcactct cagtacaatc tgctctgatg
     5881 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5941 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6001 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     6061 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6121 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6181 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6241 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6301 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6361 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6421 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6481 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6541 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6601 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6661 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6721 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6781 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6841 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6901 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6961 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7021 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7081 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7141 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7201 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7261 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7321 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7381 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7441 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7501 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7561 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7621 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7681 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7741 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7801 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7861 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac
     7921 agatcttcaa tattggccat tagccatatt attcattggt tatatagcat aaatcaatat
     7981 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
     8041 atgtccaata tgaccgccat gtt
//
