LOCUS       sequence-252646-        7982 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3303..3876
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3326..3878
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3470..3487)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3697..3716
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(3963..3991)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4586..4606
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    4770..4891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4807..4826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4861..4880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4898..5353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5035..5056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5247..5266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5380..5484
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5486..5843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5486..5677
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5509..5529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5694..5829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5878..6672
                     /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             5878..6672
                     /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(5932..5951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6542..6561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6860..6879)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6904..6951
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7280..7868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7769..7788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagtaacg
      601 gccgccagtg tgctggaatt cgcccttggg ggcggccgga tcagcgatgc cggcgggcat
      661 gacgaagcat ggctcccgct ccaccagctc gctgccgccc gagcccatgg agatcgtgcg
      721 cagcaaggcg tgctctcggc gggtccgcct caacgtcggg gggctggcgc acgaggtact
      781 ctggcgtacc ctggaccgcc tgccccgcac gcggctgggc aagctccgcg actgcaacac
      841 gcacgactcg ctgctcgagg tgtgcgatga ctacagcctc gacgacaacg agtacttctt
      901 tgaccgccac ccgggcgcct tcacctccat cctcaacttc taccgcactg ggcgactgca
      961 catgatggag gagatgtgcg cgctcagctt cagccaagag ctcgactact ggggcatcga
     1021 cgagatctac ctggagtcct gctgccaggc ccgctaccac cagaagaaag agcagatgaa
     1081 cgaggagctc aagcgtgagg ccgagaccct acgggagcgg gaaggcgagg agttcgataa
     1141 cacgtgctgc gcagagaaga ggaaaaaact ctgggaccta ctggagaagc ccaattcctc
     1201 tgtggctgcc aagatccttg ccataatttc catcatgttc atcgtcctct ccaccattgc
     1261 cctgtccctc aacacgctgc ctgagctaca gagcctcgat gagttcggcc agtccacaga
     1321 caacccccag ctggcccacg tggaggccgt gtgcatcgca tggttcacca tggagtacct
     1381 gctgaggttc ctctcctcgc ccaagaagtg gaagttcttc aagggcccac tcaatgccat
     1441 tgacttgttg gccattctgc catactatgt caccattttc ctcaccgaat ccaacaagag
     1501 cgtgctgcaa ttccagaatg tccgccgcgt ggtccagatc ttccgcatca tgcgaattct
     1561 ccgcatcctt aagcttgcac gccactccac tggcctccag tctctgggct tcactttgcg
     1621 gaggagctac aatgagttgg gcttgctcat cctcttcctt gccatgggca ttatgatctt
     1681 ctccagcctt gtcttctttg ctgagaagga tgaggacgac accaagttca aaagcatccc
     1741 agcctctttc tggtgggcca ccatcaccat gactactgtt gggtatggag acatctaccc
     1801 caagactctc ctggggaaaa ttgttggggg actctgctgc attgcaggag tcctggtgat
     1861 tgctcttccc atccccatca tcgtcaataa cttctctgag ttctataagg agcagaagag
     1921 acaggagaaa gcaatcaaac ggcgagaggc tctggagaga gccaagagga atggcagcat
     1981 cgtatccatg aacatgaagg atgcttttgc ccggagcatt gagatgatgg acattgtggt
     2041 tgagaaaaat ggggagaata tgggtaagaa agacaaagta caagataacc acttgtctcc
     2101 taacaaatgg aaatggacaa agaggacact gtctgaaacc agctcaagta agtcctttga
     2161 aaccaaggaa cagggatccc ctgaaaaagc cagatcgtct tctagtcctc agcacctgaa
     2221 cgttcagcag ttggaagaca tgtacaataa gatggccaag acccaatccc aacccatcct
     2281 caataccaag gagtcagcag cacagagcaa accaaaggaa gaacttgaaa tggagagtat
     2341 ccccagcccc gtagcccctc tgcccactcg cacagaaggg gtcattgaca tgcgaagtat
     2401 gtcaagcatt gatagtttca ttagctgtgc cacagacttc cctgaggcca ccagattctc
     2461 ccacagccct ttgacatcac tccccagcaa gactgggggc agcacagccc cagaagtggg
     2521 ctggcgggga gctctgggtg ccagtggtgg taggtttgtg gaggccaacc ccagccctga
     2581 tgccagccag cactctagtt tcttcatcga gagccccaag agttccatga aaactaacaa
     2641 ccctttgaag ctccgagcac ttaaagtcaa cttcatggag ggtgacccca gtccactcct
     2701 ccccgttcta gggatgtacc atgaccctct caggaaccgg gggagtgccg cggctgctgt
     2761 cgctggactg gagtgtgcca cgcttttgga caaggctgtg ctgagcccag agtcctccat
     2821 ctacaccaca gcaagtgcta agacaccccc ccggtctcct gagaaacaca cagcaatagc
     2881 gttcaacttt gaggcgggtg tccaccagta cattgacgca gacacagatg atgagggaca
     2941 gctgctctac agtgtggact ccagcccccc caaaagcctc cctgggagca ccagtccgaa
     3001 gttcagcacg gggacaagat cggagaaaaa ccactttgaa agctcccctt tacccacctc
     3061 ccctaagttc ttaaggcaga actgtattta ctccacagaa gcattgactg gaaaaggccc
     3121 cagtggtcag gaaaagtgca aacttgagaa ccacatctcc cctgacgtcc gtgtgttgcc
     3181 agggggagga gcccatggaa gcacacgaga tcagagcatc tgaactgccc tgcctaaggg
     3241 cgaattctgc agatatccat cacactggcg gccgctcgac ggtaccgcgg gcccgggatc
     3301 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     3361 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     3421 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     3481 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     3541 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     3601 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     3661 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     3721 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     3781 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     3841 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacctcgc gcgcctgcag
     3901 gtcgacacta gtggatccaa ggagatataa caatgagtaa aggagaagaa cttttcactg
     3961 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     4021 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     4081 ctggaaaact acctgttcca tggccaacac ttgtcactac tttcacttat ggtgttcaat
     4141 gcttttcaag atacccagat catatgaagc ggcacgactt cttcaagagc gccatgcctg
     4201 agggatacgt gcaggagagg accatctctt tcaaggacga cgggaactac aagacacgtg
     4261 ctgaagtcaa gtttgaggga gacaccctcg tcaacaggat cgagcttaag ggaatcgatt
     4321 tcaaggagga cggaaacatc ctcggccaca agttggaata caactacaac tcccacaacg
     4381 tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca
     4441 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     4501 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     4561 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     4621 cacatggcat ggatgaacta tacaaataac ggccgcgact ctagatcata atcagccata
     4681 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4741 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4801 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4861 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     4921 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4981 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5041 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5101 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5161 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5221 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5281 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5341 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5401 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5461 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     5521 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5581 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5641 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5701 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5761 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5821 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     5881 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5941 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6001 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     6061 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6121 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6181 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6241 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6301 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6361 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     6421 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6481 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6541 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6601 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6661 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6721 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6781 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6841 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     6901 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     6961 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     7021 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     7081 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     7141 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7201 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7261 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7321 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7381 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     7441 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7501 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7561 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7621 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7681 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7741 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7801 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7861 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7921 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     7981 at
//
