LOCUS       sequence-260090-        6089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for NaVSp1 bacterial sodium 
            channel. N-terminal HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6089)
  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 6089)
  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: HA tag NaVSp1 FL pIRES
FEATURES             Location/Qualifiers
     source          1..6089
                     /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"
     CDS             606..632
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(1448..2022,2023..2025,2026..2034)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1615..1632)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1842..1861
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2035..2037,2038..2040,2041..2754)
                     /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(2035..2037,2038..2040,2041..2751)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2080..2101)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2341..2360)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2688..2709
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2877..2998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2914..2933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2968..2987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3005..3460)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3142..3163)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3354..3373
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3487..3591
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3593..3950
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3593..3784
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3616..3636)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3801..3936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3985..4779
                     /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             3985..4779
                     /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(4039..4058)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4649..4668
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4967..4986)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5011..5058
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5387..5975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5876..5895
                     /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 gctagcacca
      601 tggcatatcc atatgatgtt ccagattatg ctatgcaaag aatgcaggcc tttatcgacc
      661 ggcccggttt cggccggttc attaccgccg tgatcatgct gaacgccgtc gtgctgggga
      721 tggagacgtc gccctatctg atggagcggt ggggcgggct gatcctggcc gctgaccgta
      781 tctgcctagg catcttcatc gtcgagatcg cgctgaaact gacggcgcag cggcagcgct
      841 ttttcctgaa cggctggaac ctgttcgatt tcgtgatcgt cgggatcgcc ctgatccccg
      901 ccgccggggg tctgtcggtg ctgcgggcgc tgcgcatcct gcgggtgctt cgcgtgatct
      961 cggtcgcgcc gcgcttgcgc cgcgtggtcg aaggcttcat cactgcgctg cccggcatgg
     1021 ccagcgtgtt tctgttgatg accatcatct tctacatcgg cgcggtgatc gcgaccaagc
     1081 tgttcgccgc ctcctttccc gactggttcg gcgatctggg tctgtcggcc tatacgctgt
     1141 tccagatcat gacgctggaa agctggtcga tgggcatcgt tcgcccggtg atgcaggtct
     1201 atccctatgc ctggctgttc tttgtcccct tcatcatgat caccacattt gcggtggtta
     1261 acctgctggt cggtctgatc gtgaactcga tgcaggacgc tcatcacgcc gaggatggcg
     1321 agcggaccga tgcctatcgg gacgaggttc tggcccgcct ggaacagatc gaccagcggc
     1381 tgaatgcgct tggtgaaacc aaaaagtgag aattctgcag tcgacggtac cgcgggcccg
     1441 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     1501 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     1561 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     1621 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     1681 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     1741 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     1801 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     1861 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     1921 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     1981 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     2041 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2101 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2161 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2221 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2281 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2341 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2401 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2461 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2521 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2581 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2641 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2701 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     2761 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2821 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2881 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2941 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     3001 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3061 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3121 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3181 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3241 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3301 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3361 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3421 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3481 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3541 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3601 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3661 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3721 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3781 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3841 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3901 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3961 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4021 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4081 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4141 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4201 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4261 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4321 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4381 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4441 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4501 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4561 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4621 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4681 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4741 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4801 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4861 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4921 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4981 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5041 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5101 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5161 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5221 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5281 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5341 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5521 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6001 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6061 ctgtggataa ccgtattacc gccatgcat
//
