LOCUS       sequence-260247-        7982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Kv2.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7982)
  AUTHORS   Kang SK, Vanoye CG, Misra SN, Echevarria DM, Calhoun JD, O'Connor 
            JB, Fabre KL, McKnight D, Demmer L, Goldenberg P, Grote LE, 
            Thiffault I, Saunders C, Strauss KA, Torkamani A, van der Smagt J, 
            van Gassen K, Carson RP, Diaz J, Leon E, Jacher JE, Hannibal MC, 
            Litwin J, Friedman NR, Schreiber A, Lynch B, Poduri A, Marsh ED, 
            Goldberg EM, Millichap JJ, George AL Jr, Kearney JA
  TITLE     Spectrum of KV 2.1 Dysfunction in KCNB1-Associated 
            Neurodevelopmental Disorders.
  JOURNAL   Ann Neurol. 2019 Dec;86(6):899-912. doi: 10.1002/ana.25607. Epub 
            2019 Oct 24.
  PUBMED    31600826
REFERENCE   2  (bases 1 to 7982)
  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: "10.1002/ana.25607";
            journalName: "Ann Neurol"; date: "2019-12"; volume: "86"; issue: 
            "6"; pages: "899-912"
COMMENT     Sequence Label: pIRES2-smGFP-Kv2.1
FEATURES             Location/Qualifiers
     source          1..7982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2939..3512
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2962..3514
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3106..3123)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3333..3352
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(3599..3627)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4222..4242
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    4406..4527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4443..4462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4497..4516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4534..4989)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4671..4692)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4883..4902
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5016..5120
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5122..5479
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5122..5313
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5145..5165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5330..5465
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5514..6308
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5514..6308
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5568..5587)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6178..6197
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6496..6515)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6540..6587
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6916..7504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7405..7424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7679..7982
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gtaacggccg
      241 ccagtgtgct ggaattcgcc cttgggggcg gccggatcag cgatgccggc gggcatgacg
      301 aagcatggct cccgctccac cagctcgctg ccgcccgagc ccatggagat cgtgcgcagc
      361 aaggcgtgct ctcggcgggt ccgcctcaac gtcggggggc tggcgcacga ggtactctgg
      421 cgtaccctgg accgcctgcc ccgcacgcgg ctgggcaagc tccgcgactg caacacgcac
      481 gactcgctgc tcgaggtgtg cgatgactac agcctcgacg acaacgagta cttctttgac
      541 cgccacccgg gcgccttcac ctccatcctc aacttctacc gcactgggcg actgcacatg
      601 atggaggaga tgtgcgcgct cagcttcagc caagagctcg actactgggg catcgacgag
      661 atctacctgg agtcctgctg ccaggcccgc taccaccaga agaaagagca gatgaacgag
      721 gagctcaagc gtgaggccga gaccctacgg gagcgggaag gcgaggagtt cgataacacg
      781 tgctgcgcag agaagaggaa aaaactctgg gacctactgg agaagcccaa ttcctctgtg
      841 gctgccaaga tccttgccat aatttccatc atgttcatcg tcctctccac cattgccctg
      901 tccctcaaca cgctgcctga gctacagagc ctcgatgagt tcggccagtc cacagacaac
      961 ccccagctgg cccacgtgga ggccgtgtgc atcgcatggt tcaccatgga gtacctgctg
     1021 aggttcctct cctcgcccaa gaagtggaag ttcttcaagg gcccactcaa tgccattgac
     1081 ttgttggcca ttctgccata ctatgtcacc attttcctca ccgaatccaa caagagcgtg
     1141 ctgcaattcc agaatgtccg ccgcgtggtc cagatcttcc gcatcatgcg aattctccgc
     1201 atccttaagc ttgcacgcca ctccactggc ctccagtctc tgggcttcac tttgcggagg
     1261 agctacaatg agttgggctt gctcatcctc ttccttgcca tgggcattat gatcttctcc
     1321 agccttgtct tctttgctga gaaggatgag gacgacacca agttcaaaag catcccagcc
     1381 tctttctggt gggccaccat caccatgact actgttgggt atggagacat ctaccccaag
     1441 actctcctgg ggaaaattgt tgggggactc tgctgcattg caggagtcct ggtgattgct
     1501 cttcccatcc ccatcatcgt caataacttc tctgagttct ataaggagca gaagagacag
     1561 gagaaagcaa tcaaacggcg agaggctctg gagagagcca agaggaatgg cagcatcgta
     1621 tccatgaaca tgaaggatgc ttttgcccgg agcattgaga tgatggacat tgtggttgag
     1681 aaaaatgggg agaatatggg taagaaagac aaagtacaag ataaccactt gtctcctaac
     1741 aaatggaaat ggacaaagag gacactgtct gaaaccagct caagtaagtc ctttgaaacc
     1801 aaggaacagg gatcccctga aaaagccaga tcgtcttcta gtcctcagca cctgaacgtt
     1861 cagcagttgg aagacatgta caataagatg gccaagaccc aatcccaacc catcctcaat
     1921 accaaggagt cagcagcaca gagcaaacca aaggaagaac ttgaaatgga gagtatcccc
     1981 agccccgtag cccctctgcc cactcgcaca gaaggggtca ttgacatgcg aagtatgtca
     2041 agcattgata gtttcattag ctgtgccaca gacttccctg aggccaccag attctcccac
     2101 agccctttga catcactccc cagcaagact gggggcagca cagccccaga agtgggctgg
     2161 cggggagctc tgggtgccag tggtggtagg tttgtggagg ccaaccccag ccctgatgcc
     2221 agccagcact ctagtttctt catcgagagc cccaagagtt ccatgaaaac taacaaccct
     2281 ttgaagctcc gagcacttaa agtcaacttc atggagggtg accccagtcc actcctcccc
     2341 gttctaggga tgtaccatga ccctctcagg aaccggggga gtgccgcggc tgctgtcgct
     2401 ggactggagt gtgccacgct tttggacaag gctgtgctga gcccagagtc ctccatctac
     2461 accacagcaa gtgctaagac acccccccgg tctcctgaga aacacacagc aatagcgttc
     2521 aactttgagg cgggtgtcca ccagtacatt gacgcagaca cagatgatga gggacagctg
     2581 ctctacagtg tggactccag cccccccaaa agcctccctg ggagcaccag tccgaagttc
     2641 agcacgggga caagatcgga gaaaaaccac tttgaaagct cccctttacc cacctcccct
     2701 aagttcttaa ggcagaactg tatttactcc acagaagcat tgactggaaa aggccccagt
     2761 ggtcaggaaa agtgcaaact tgagaaccac atctcccctg acgtccgtgt gttgccaggg
     2821 ggaggagccc atggaagcac acgagatcag agcatctgaa ctgccctgcc taagggcgaa
     2881 ttctgcagat atccatcaca ctggcggccg ctcgacggta ccgcgggccc gggatccgcc
     2941 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     3001 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     3061 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     3121 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     3181 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     3241 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     3301 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     3361 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca
     3421 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     3481 acgtggtttt cctttgaaaa acacgatgat aatatggcca cctcgcgcgc ctgcaggtcg
     3541 acactagtgg atccaaggag atataacaat gagtaaagga gaagaacttt tcactggagt
     3601 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     3661 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     3721 aaaactacct gttccatggc caacacttgt cactactttc acttatggtg ttcaatgctt
     3781 ttcaagatac ccagatcata tgaagcggca cgacttcttc aagagcgcca tgcctgaggg
     3841 atacgtgcag gagaggacca tctctttcaa ggacgacggg aactacaaga cacgtgctga
     3901 agtcaagttt gagggagaca ccctcgtcaa caggatcgag cttaagggaa tcgatttcaa
     3961 ggaggacgga aacatcctcg gccacaagtt ggaatacaac tacaactccc acaacgtata
     4021 catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta gacacaacat
     4081 tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     4141 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
     4201 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     4261 tggcatggat gaactataca aataacggcc gcgactctag atcataatca gccataccac
     4321 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4381 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4441 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4501 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     4561 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4621 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4681 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4741 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4801 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4861 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4921 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4981 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5041 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5101 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     5161 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5221 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5281 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5341 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5401 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5461 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     5521 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5581 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5641 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5701 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5761 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5821 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5881 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5941 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6001 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     6061 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6121 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6181 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6241 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6301 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6361 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6421 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6481 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     6541 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6601 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6661 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6721 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6781 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7021 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     7621 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     7681 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7741 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7801 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     7861 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     7921 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7981 tg
//
