LOCUS       sequence-260100-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for (NaVSp1 1-236 NaVAe1 265-288) 
            chimera bacterial sodium channel..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  AUTHORS   Arrigoni C, Rohaim A, Shaya D, Findeisen F, Stein RA, Nurva SR, 
            Mishra S, Mchaourab HS, Minor DL Jr
  TITLE     Unfolding of a Temperature-Sensitive Domain Controls Voltage-Gated 
            Channel Activation.
  JOURNAL   Cell. 2016 Feb 25;164(5):922-36. doi: 10.1016/j.cell.2016.02.001.
  PUBMED    26919429
REFERENCE   2  (bases 1 to 6084)
  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.cell.2016.02"; journalName: "Cell"; date: "2016-02-25- 
            25"; volume: "164"; issue: "5"; pages: "922-36"
COMMENT     Sequence Label: NaVSp1 Ae1 CC pIRES
FEATURES             Location/Qualifiers
     source          1..6084
                     /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    join(1443..2017,2018..2020,2021..2029)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1610..1627)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1837..1856
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2030..2032,2033..2035,2036..2749)
                     /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(2030..2032,2033..2035,2036..2746)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2075..2096)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2336..2355)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2683..2704
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2872..2993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2909..2928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2963..2982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3000..3455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3349..3368
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3482..3586
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3588..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3588..3779
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3980..4774
                     /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             3980..4774
                     /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(4034..4053)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4644..4663
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4962..4981)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5006..5053
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5382..5970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5871..5890
                     /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 gctagcgcta
      601 ccggactcag atctcgagat gcaaagaatg caggccttta tcgaccggcc cggtttcggc
      661 cggttcatta ccgccgtgat catgctgaac gccgtcgtgc tggggatgga gacgtcgccc
      721 tatctgatgg agcggtgggg cgggctgatc ctggccgctg accgtatctg cctaggcatc
      781 ttcatcgtcg agatcgcgct gaaactgacg gcgcagcggc agcgcttttt cctgaacggc
      841 tggaacctgt tcgatttcgt gatcgtcggg atcgccctga tccccgccgc cgggggtctg
      901 tcggtgctgc gggcgctgcg catcctgcgg gtgcttcgcg tgatctcggt cgcgccgcgc
      961 ttgcgccgcg tggtcgaagg cttcatcact gcgctgcccg gcatggccag cgtgtttctg
     1021 ttgatgacca tcatcttcta catcggcgcg gtgatcgcga ccaagctgtt cgccgcctcc
     1081 tttcccgact ggttcggcga tctgggtctg tcggcctata cgctgttcca gatcatgacg
     1141 ctggaaagct ggtcgatggg catcgttcgc ccggtgatgc aggtctatcc ctatgcctgg
     1201 ctgttctttg tccccttcat catgatcacc acatttgcgg tggttaacct gctggtcggt
     1261 ctgatcgtga actcgatgca ggacgctcat cacgccgagg atggcgagcg gaccgatgcc
     1321 tatcggctgg agatgctgca gttgatccgc gatctcagca gcaaggtcga ccggctggaa
     1381 cgtcgctccg gcaagcggta ataagaattc tgcagtcgac ggtaccgcgg gcccgggatc
     1441 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     1501 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     1561 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     1621 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     1681 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     1741 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     1801 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     1861 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     1921 tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     1981 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa
     2041 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2101 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2161 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2221 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2281 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2341 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2401 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2461 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2521 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2581 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2641 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2701 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     2761 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2821 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3001 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3061 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3121 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3181 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3241 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3301 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3361 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3421 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3601 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3961 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4021 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4081 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4141 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4201 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4261 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4321 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4381 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4441 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4501 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4561 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4621 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4681 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4741 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4801 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4861 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4921 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4981 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5041 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5101 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5161 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5221 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5281 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5521 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5761 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5821 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5881 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5941 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6001 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6061 gataaccgta ttaccgccat gcat
//
