LOCUS       sequence-167489-        8486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression Kv10.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8486)
  AUTHORS   Chen Y, Sanchez A, Rubio ME, Kohl T, Pardo LA, Stuhmer W
  TITLE     Functional K(v)10.1 channels localize to the inner nuclear membrane.
  JOURNAL   PLoS One. 2011 May 3;6(5):e19257. doi: 10.1371/journal.pone.0019257.
  PUBMED    21559285
REFERENCE   2  (bases 1 to 8486)
  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"; journalName: "PLoS One.";
            date: "2011-05-3"; volume: "6(5):e19257. doi"; pages: " 
            10.1371/journal.pone.0019257"
COMMENT     Sequence Label: pcKv10.1
FEATURES             Location/Qualifiers
     source          1..8486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2680..2694)
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     primer_bind     complement(3987..4003)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3987..4003)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4038..4056)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4039..4056)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4076..4093)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4082..4306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4352..4780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4439..4458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4649..4670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4789..4809)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4794..5124
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4975..5110
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5037..5056
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5191..5985
                     /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(5245..5264)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5855..5874
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6159..6280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6196..6215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6250..6269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6329..6345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6329..6345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6342..6364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6353..6369
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6377..6383,6384..6401,6402..6407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6422..6443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6560..6577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6731..7319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6811..6830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7490..8281,8282..8350))
                     /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     8113..8132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8351..8455)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca
      961 ctggcggccg gaattccggg cccgccggac cccgagctgc tgggaggatg accatggctg
     1021 ggggcaggag gggactagtg gcccctcaaa acacgtttct ggagaatatt gttcggcggt
     1081 ccaatgatac taattttgtg ttggggaatg ctcagatagt ggactggcct attgtgtaca
     1141 gcaatgatgg attttgcaag ctgtctggct atcacagggc agaagtgatg caaaaaagca
     1201 gcacctgcag ttttatgtat ggggagctga ctgataaaga cacgattgaa aaagtgcggc
     1261 aaacatttga gaactatgag atgaattcct ttgaaattct gatgtacaag aagaacagga
     1321 cacctgtgtg gttctttgtg aaaattgctc caattcgaaa cgaacaggat aaagtggttt
     1381 tatttctttg cactttcagt gacataacag ctttcaaaca gccaattgag gatgattcat
     1441 gtaaaggctg ggggaagttt gctcggctga caagagcact gacaagcagc aggggtgtcc
     1501 tgcagcagct ggctccaagc gtgcaaaaag gcgagaatgt ccacaagcac tcccgcctgg
     1561 cagaggtcct acagctgggc tcagacatcc ttccccagta caagcaagag gcaccaaaga
     1621 ctccccctca catcatctta cattattgtg tttttaagac cacgtgggat tggatcatct
     1681 tgatcttgac cttctataca gccatcttgg tcccttataa tgtctccttc aaaaccaggc
     1741 agaataatgt ggcctggctg gttgttgata gcatcgtgga tgttatcttt ttggtggaca
     1801 ttgtgctcaa ttttcatacc acctttgttg gaccagcagg ggaggtgatt tctgacccca
     1861 aacttatccg catgaactac ctgaagacgt ggtttgtgat tgaccttctg tcctgtttgc
     1921 catatgatgt catcaacgct tttgagaacg tggatgaggg catcagcagc ctgttcagct
     1981 ctctaaaagt tgtccggctg ctccgtcttg ggcgagtggc ccgtaagctg gaccactaca
     2041 ttgaatatgg agctgctgtg ctggtcctgc tggtgtgtgt gtttgggctg gctgcacact
     2101 ggatggcctg catctggtac agcattgggg actatgagat ctttgacgag gacaccaaga
     2161 caatccgcaa caacagctgg ctgtaccaac tagcgatgga cattggcacc ccttaccagt
     2221 ttaatgggtc tggctcaggg aagtgggaag gtggtcccag caagaattct gtctacatct
     2281 cctcgttgta tttcacaatg accagcctca ccagtgtggg ctttgggaac atcgccccat
     2341 ccacagacat tgagaagatc tttgcagtgg ccatcatgat gattggctca cttctctatg
     2401 ccaccatctt cgggaatgtg acgactattt tccaacagat gtatgccaac accaacagat
     2461 accatgagat gctcaacagt gttcgggact tcctgaagct ctaccaggtg ccaaaaggat
     2521 tgagtgagcg agtaatggat tatattgtgt ccacttggtc catgtccaga ggcattgaca
     2581 cagagaaggt cctgcagatc tgccccaagg acatgagagc cgacatctgc gtgcacctga
     2641 accgcaaggt gttcaaggag cacccggcct tccggctggc cagtgatggc tgcctccggg
     2701 cactggccat ggagttccag acggtgcact gtgccccagg ggacctcatc taccatgcag
     2761 gagagagcgt tgacagcctc tgctttgtgg tttctggctc cctggaggtg atccaagatg
     2821 atgaggtggt ggccattcta ggaaaaggag acgtgtttgg agatgtgttc tggaaggaag
     2881 ccacccttgc ccagtcctgt gccaatgtta gggccttgac ctactgtgat ctgcatgtga
     2941 tcaagcggga tgccctgcag aaagtgctgg aattctacac ggccttctcc cattccttct
     3001 cccggaacct gattctgacg tacaacttga ggaagaggat tgtgttccgg aagatcagcg
     3061 atgtgaaacg tgaagaggaa gaacgcatga aacgaaagaa tgaggccccc ctgatcttgc
     3121 ccccggacca ccctgtccgg cgcctcttcc agagattccg acagcagaaa gaggccaggc
     3181 tggcagctga gagagggggc cgggacctgg atgacctaga tgtggagaag ggcaatgtcc
     3241 ttacagagca tgcctccgcc aaccacagcc tcgtgaaggc cagcgtggtc accgtgcgtg
     3301 agagtcctgc cacgcccgta tccttccagg cagcctccac ctccggggtg ccagaccacg
     3361 caaagctaca ggcgccaggg tccgagtgcc tgggccccaa ggggggcggg ggcgattgtg
     3421 ccaagcgcaa aagctgggcc cgcttcaaag atgcttgcgg gaagagtgag gactggaaca
     3481 aggtgtccaa ggctgagtcg atggagacac ttcccgagag gacaaaagcg tcaggcgagg
     3541 ccacactgaa gaagacagac tcgtgtgaca gtggcatcac caagagcgac ttgcgcctgg
     3601 acaacgtggg tgaggccagg agtccccagg atcggagtcc catcctggca gaggtcaagc
     3661 attcgttcta ccccatccct gagcagacgc tgcaggccac agtcctggag gtgaggcacg
     3721 agctgaagga ggacatcaag gccttaaacg ccaaaatgac caatattgag aaacagctct
     3781 ctgagatact caggatatta acttccagaa gatcctctca gtctcctcag gagttgtttg
     3841 aaatatcgag gccacagtcc ccagaatcag agagagacat ttttggagcc agctgagagg
     3901 tctatttaaa aaaaaagtca gagacagata cctccaaccc tgccgtcacc accaccccta
     3961 ccacccggaa ttcctgcagc ccgggggatc cactagttct agagcggccg ctcgagcatg
     4021 catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
     4081 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4141 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4201 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4261 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4321 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4381 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4441 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4501 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4561 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4621 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4681 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     4741 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4801 agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat
     4861 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4921 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4981 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5041 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     5101 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     5161 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5221 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5281 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5341 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5401 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5461 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5521 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5581 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5641 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5701 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5761 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5821 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5881 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5941 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6001 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6061 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6121 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6181 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6241 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     6301 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6361 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6421 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6481 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6541 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6601 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6901 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     6961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7141 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7441 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7621 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7801 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8101 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8341 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8461 ttccccgaaa agtgccacct gacgtc
//
