LOCUS       sequence-217427-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses worm KVS-1 C113S in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  AUTHORS   Sesti F, Liu S, Cai SQ
  TITLE     Oxidation of potassium channels by ROS: a general mechanism of aging
            and neurodegeneration?
  JOURNAL   Trends Cell Biol. 2010 Jan;20(1):45-51. doi: 
            10.1016/j.tcb.2009.09.008. Epub 2009 Oct 21.
  PUBMED    19850480
REFERENCE   2  (bases 1 to 6955)
  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: 
            "10.1016/j.tcb.2009.09.008"; journalName: "Trends Cell Biol"; date: 
            "2010-01"; volume: "20"; issue: "1"; pages: "45-51"
COMMENT     Sequence Label: C113S-HA
FEATURES             Location/Qualifiers
     source          1..6955
                     /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             2430..2456
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2522..2643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2533..2552)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2587..2606
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2829..3284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2916..2935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3126..3147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3401..3758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3401..3592
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3424..3444)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3609..3744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3809..4603
                     /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(3863..3882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4473..4492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4667..4715
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4756..4775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4875..4897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4935..4953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5021..5125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5126..5194,5195..5986)
                     /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(5344..5363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6157..6745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6646..6665
                     /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 tgagcacgga aaggctgatg tatctcgatt taaataaagg caaccaaatt gccatggtga
     1021 ttcccagcac caggaaagga catgctgcat ctcgtgacaa ttttggcaac tctaaccatc
     1081 atggtgacga gattgtgcac cgactcaaca atgggagacg ttcagagata gatgaagtaa
     1141 agcccgacct tgtgtcaggg gccggcgaca ctgtgatcaa gttgaatgtt ggagggacat
     1201 cattcagaat aaggatatcc agcatctttg caagatcggc ggacgagaaa ctgtccgcgt
     1261 ttgcacaatt aaaccatgag cagagacttc aagcatctga cgcgtttctt atgccacaag
     1321 aggagtacta cttggaaagg tcggctgccc tttttgattc gatcttcaaa tactacatat
     1381 ctggccaact acacagaccg cttgatgtat gccccacaga atttgccacc gaactcgcct
     1441 actggaagat aaacgacggg ctcacgtcaa actgctgttg gaggggatac aatatgagtc
     1501 cttcactaga agaactggtg gccccgaaag agaaaaaaat agagctgcac gaccgttcaa
     1561 tgcgctcacg gattcatcaa ttctgcgaag gggacggctc acttttctcg actttgttta
     1621 cttttggcag catcactttt gttcttatta gtgttattgg actggttttg ggatccattc
     1681 acgaatttca agtacctatc tataaaaaag gaggaaaatc agagaggatc agcaactgga
     1741 gtatcgtgga aaacgaaaac attatatggg agccacatcc gatttttggg tatgtggaga
     1801 cagtgtgtat catctggttt actgtagaat tcatcgtacg gttagcagtt acaccaaata
     1861 gattccaatt tttggtggga atcatgaata ttgtcgatat gatcgctatt attccattct
     1921 atttggaact cgggttggcg ttgtttggaa ttgatgtcgc tagtctgagc gacattaaag
     1981 gagcattatt agttgttaga gtgctccgtg tgcctcgtgt tgtgagaatt ttgaaacttg
     2041 gaagatactc atctggtatg cgaacattcg ctttgacact gagaagttcc gctcgccagc
     2101 tgggaatgat gggaatggtc ctgtcaaccg gtgtcgtatt cttctcaaca cttttatact
     2161 tcgtcgagaa agacgaaaaa gacacgccgt tcacttcgat tccagctgct ttttggtggg
     2221 caatcgtcac gatgacaact gttggatatg gagattatgt tcctgtgaca gttcctggaa
     2281 aactgattgc aagtggagca attatcagtg gagtattggt acttgctctt cctatcacta
     2341 tcatcgttga caactttatg aaagtttctg gaaatgtgaa tgccaacatc ttctcatctg
     2401 caaaccaacg acctattgat gtggcagaat acccatacga cgtcccagac tacgcttaat
     2461 ctagagtcga cccgggcggc cgcttccctt tagtgagggt taatgcttcg agcagacatg
     2521 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2581 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2641 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2701 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc gatccgggct
     2761 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2821 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2881 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2941 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3001 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3061 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3121 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3181 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3241 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     3301 ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc
     3361 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc
     3421 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3481 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3601 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3661 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3721 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa
     3781 cagtctcgaa cttaaggcta gagccaccat gattgaacaa gatggattgc acgcaggttc
     3841 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3901 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3961 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4021 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4081 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4141 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4201 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4261 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4321 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4381 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4441 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4501 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4561 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4621 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     4681 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt
     4741 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4801 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4861 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4921 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4981 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5101 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5161 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5221 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5281 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5341 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5401 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5461 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5521 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5581 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5641 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5701 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5761 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5821 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5881 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5941 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6001 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6061 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6301 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6361 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6421 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6481 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6541 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6601 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6661 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6721 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6781 cttttgctgg ccttttgctc acatggctcg acagatcttc aatattggcc attagccata
     6841 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     6901 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
