LOCUS       sequence-342918-        5696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of KCNE1 and creating variants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5696)
  AUTHORS   Kuenze G, Vanoye CG, Desai RR, Adusumilli S, Brewer KR, Woods H, 
            McDonald EF, Sanders CR, George AL Jr, Meiler J
  TITLE     Allosteric mechanism for KCNE1 modulation of KCNQ1 potassium channel
            activation.
  JOURNAL   Elife. 2020 Oct 23;9. pii: 57680. doi: 10.7554/eLife.57680.
  PUBMED    33095155
REFERENCE   2  (bases 1 to 5696)
  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: "Elife."; 
            date: "2020-10-23"; volume: "9. pii: 57680. doi"; pages: " 
            10.7554/eLife.57680"
COMMENT     Sequence Label: pIRES_EGFP_GA_KCNE1
FEATURES             Location/Qualifiers
     source          1..5696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(450..1024,1025..1027,1028..1036)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(617..634)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     844..863
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1037..1039,1040..1042,1043..1756)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1037..1039,1040..1042,1043..1753)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1082..1103)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1343..1362)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1690..1711
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1879..2000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1916..1935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1970..1989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2007..2462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2144..2165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2356..2375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2489..2593
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2595..2952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2595..2786
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2618..2638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2803..2938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2987..3781
                     /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             2987..3781
                     /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(3041..3060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3651..3670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3969..3988)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4013..4060
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4389..4977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4878..4897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5152..5455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5456..5659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5610..5630
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atgatcctgt ctaacaccac agcggtgacg ccctttctga ccaagctgtg gcaggagaca
       61 gttcagcagg gtggcaacat gtcgggcctg gcccgcaggt ccccccgcag cggtgacggc
      121 aagctggagg ccctctacgt cctcatggta ctgggattct tcggcttctt caccctgggc
      181 atcatgctga gctacatccg ctccaagaag ctggagcact cgaacgaccc attcaacgtc
      241 tacatcgagt ccgatgcctg gcaagagaag gacaaggcct atgtccaggc ccgggtcctg
      301 gagagctaca ggtcgtgcta tgtcgttgaa aaccatctgg ccatagaaca acccaacaca
      361 caccttcctg agacgaagcc ttccccatga ggatccaccg gtcatccacg cctgagatct
      421 agacaaactt cctagactgc atggatccgc ccctctccct cccccccccc taacgttact
      481 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
      541 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
      601 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
      661 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
      721 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
      781 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
      841 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
      901 tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta gtcgaggtta
      961 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     1021 taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1081 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1141 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1201 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1261 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1321 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1381 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1441 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1501 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1561 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1621 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1681 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1741 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1801 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1861 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1921 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1981 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2041 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2101 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2221 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2461 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2521 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2581 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2641 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2701 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2761 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2821 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2881 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2941 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3001 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3061 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3121 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3181 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3241 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3301 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3361 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3421 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3481 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3541 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3601 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3661 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3721 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3781 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3841 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3901 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3961 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4021 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4081 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4141 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4201 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4261 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4321 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4381 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4441 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4501 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4561 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4621 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4681 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4741 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4801 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4861 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4921 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4981 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5041 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     5101 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5161 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5221 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5281 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     5341 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     5401 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     5461 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     5521 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     5581 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     5641 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgcg gccgcc
//
