LOCUS       sequence-217435-        7235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rat Kv3.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  AUTHORS   Cotella D, Hernandez-Enriquez B, Duan Z, Wu X, Gazula VR, Brown MR, 
            Kaczmarek LK, Sesti F
  TITLE     An evolutionarily conserved mode of modulation of Shaw-like K(+) 
            channels.
  JOURNAL   FASEB J. 2013 Apr;27(4):1381-93. doi: 10.1096/fj.12-222778. Epub 
            2012 Dec 11.
  PUBMED    23233530
REFERENCE   2  (bases 1 to 7235)
  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.12-222778"; journalName: "FASEB J"; date: "2013-04"; 
            volume: "27"; issue: "4"; pages: "1381-93"
COMMENT     Sequence Label: rat Kv3.1-HA
FEATURES             Location/Qualifiers
     source          1..7235
                     /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"
     regulatory      579..588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2346..2372
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2422..2543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2433..2452)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2487..2506
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2729..3184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2816..2835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3026..3047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3301..3658
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3301..3492
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3324..3344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3509..3644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3709..4503
                     /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(3763..3782)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4373..4392
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4567..4615
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4656..4675)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4775..4797
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4835..4853)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4921..5025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5026..5094,5095..5886)
                     /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(5244..5263)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6057..6645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6546..6565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6856..7235
                     /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 agcctcgagc caccatgggc caaggggacg
      601 agagcgagcg catcgtgatc aacgtgggcg gcacgcgcca ccagacgtac cgctcgacgc
      661 tgcgcacgct gcccggcacg cggcttgcct ggctggcgga gccggacgcc cacagccact
      721 tcgactatga cccgcgcgct gacgagttct tcttcgaccg ccacccgggc gtcttcgctc
      781 acatcctgaa ctattaccgc accggcaagt tacactgccc ggccgacgtg tgcgggccgc
      841 tctacgagga ggagctggcc ttctggggca tcgacgagac ggacgtggag ccctgctgct
      901 ggatgacgta tcgccagcac cgcgacgctg aggaggcgct ggacagcttt ggtggcgcgc
      961 ccttggacaa cagcgccgac gacgcggacg ccgacggccc cggcgactcg ggcgacggcg
     1021 aggacgagct ggagatgacc aagagattgg cgctcagtga ctccccagat ggccggcctg
     1081 gcggcttctg gcgccgctgg cagccacgca tctgggcact gttcgaggac ccctactcat
     1141 cccgctatgc gcggtacgtg gcctttgcct ccctcttctt catcctggtc tccatcacaa
     1201 ccttctgtct ggagacccac gagcgcttca accccatcgt gaacaagaca gaaattgaga
     1261 acgttcgaaa tggcacacaa gtgcggtact accgggaagc ggagacggag gccttcctca
     1321 cctacatcga gggtgtctgc gtggtctggt tcaccttcga gttcctcatg cgtgttgtct
     1381 tctgtcccaa caaggtggag ttcatcaaga actccctcaa tatcattgac ttcgtggcca
     1441 tcctcccctt ctacctggag gtgggcctaa gtggcctgtc ctccaaagct gccaaggacg
     1501 tgctaggctt cctgcgcgtc gtgcgcttcg tgcgcatcct ccgtatcttc aagctgaccc
     1561 gccattttgt gggcctgcgg gtcctgggcc acacgctccg tgccagcacc aacgagttcc
     1621 tgctgcttat caacttcctg gccctgggtg tgctcatctt tgccaccatg atctactatg
     1681 ctgagaggat aggggcacag cccaatgacc ccagcgccag cgaacacaca cactttaaaa
     1741 atatccccat cggcttctgg tgggctgtgg tcaccatgac gacactgggc tatggagaca
     1801 tgtatcctca gacgtggtct gggatgttgg tgggagcact gtgcgccctg gctggtgtgc
     1861 tgaccattgc catgccggtg cccgtcatcg tgaacaattt tgggatgtac tactctctag
     1921 ccatggctaa gcagaaacta ccaaagaaaa aaaagaagca tattccgcgg ccaccacagc
     1981 tgggatctcc caattattgt aaatctgtcg taaactctcc acaccacagt actcagagtg
     2041 acacatgtcc gctggcccag gaagaaattt tagaaattaa cagagcagat tccaaactga
     2101 atggggaggt ggcgaaggcc gcgctggcga acgaagactg cccccacata gaccaggccc
     2161 tcactcccga tgagggcctg ccctttaccc gctcgggcac ccgcgagaga tacggaccct
     2221 gcttcctctt atcaaccggg gagtacgcgt gcccgcctgg tggaggaatg agaaaggatc
     2281 tttgcaaaga aagccctgtc attgctaagt atatgccgac agaggctgtg agagtgactg
     2341 aattctaccc atacgatgtt ccagattacg cttaggcggc cgcttccctt tagtgagggt
     2401 taatgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2461 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2521 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2581 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcc
     2641 gataaggatc gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2701 acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg
     2761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2881 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2941 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3061 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3121 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3181 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg
     3241 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
     3301 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3481 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3661 gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat gattgaacaa
     3721 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3781 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3841 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3901 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3961 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4021 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4081 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4141 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4201 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4261 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4321 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4381 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4441 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4501 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     4561 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4621 atagcgataa ggatccgcgt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4681 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4741 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4801 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4861 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4921 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4981 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5041 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5101 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5161 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5221 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5281 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5341 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5401 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5461 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5521 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5581 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5641 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5701 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5761 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5821 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5881 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5941 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6001 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatcttc
     6721 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt
     6781 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     6841 tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca attacggggt
     6901 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6961 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7021 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     7081 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     7141 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     7201 agtacatcta cgtattagtc atcgctatta ccatg
//
