LOCUS       sequence-260101-        6078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for NaVSp1 bacterial sodium 
            channel..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6078)
  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 6078)
  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 FL pIRES
FEATURES             Location/Qualifiers
     source          1..6078
                     /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(1437..2011,2012..2014,2015..2023)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1604..1621)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1831..1850
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2024..2026,2027..2029,2030..2743)
                     /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(2024..2026,2027..2029,2030..2740)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2069..2090)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2330..2349)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2677..2698
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2866..2987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2903..2922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2957..2976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2994..3449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3131..3152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3343..3362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3476..3580
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3582..3939
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3582..3773
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3605..3625)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3790..3925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3974..4768
                     /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             3974..4768
                     /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(4028..4047)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4638..4657
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4956..4975)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5000..5047
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5376..5964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5865..5884
                     /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 atctcgagac catgcaaaga atgcaggcct ttatcgaccg gcccggtttc
      661 ggccggttca ttaccgccgt gatcatgctg aacgccgtcg tgctggggat ggagacgtcg
      721 ccctatctga tggagcggtg gggcgggctg atcctggccg ctgaccgtat ctgcctaggc
      781 atcttcatcg tcgagatcgc gctgaaactg acggcgcagc ggcagcgctt tttcctgaac
      841 ggctggaacc tgttcgattt cgtgatcgtc gggatcgccc tgatccccgc cgccgggggt
      901 ctgtcggtgc tgcgggcgct gcgcatcctg cgggtgcttc gcgtgatctc ggtcgcgccg
      961 cgcttgcgcc gcgtggtcga aggcttcatc actgcgctgc ccggcatggc cagcgtgttt
     1021 ctgttgatga ccatcatctt ctacatcggc gcggtgatcg cgaccaagct gttcgccgcc
     1081 tcctttcccg actggttcgg cgatctgggt ctgtcggcct atacgctgtt ccagatcatg
     1141 acgctggaaa gctggtcgat gggcatcgtt cgcccggtga tgcaggtcta tccctatgcc
     1201 tggctgttct ttgtcccctt catcatgatc accacatttg cggtggttaa cctgctggtc
     1261 ggtctgatcg tgaactcgat gcaggacgct catcacgccg aggatggcga gcggaccgat
     1321 gcctatcggg acgaggttct ggcccgcctg gaacagatcg accagcggct gaatgcgctt
     1381 ggtgaaacca aaaagtgaga attctgcagt cgacggtacc gcgggcccgg gatccgcccc
     1441 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     1501 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     1561 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     1621 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     1681 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     1741 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     1801 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     1861 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca
     1921 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     1981 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga
     2041 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2101 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2161 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2221 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2281 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2341 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2401 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2461 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2521 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2581 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     2641 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2701 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag
     2761 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2821 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3001 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3061 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3121 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3181 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3241 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3301 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3361 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3421 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3481 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3541 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3601 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3661 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3781 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3841 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3901 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3961 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4021 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4081 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4141 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4201 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4261 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4321 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4381 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4441 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4501 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4561 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4621 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4681 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4741 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4801 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4861 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4921 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4981 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5041 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5101 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5161 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5221 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5281 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5341 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5401 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5461 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5521 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5581 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5641 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5701 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5761 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5821 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5881 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5941 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6001 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6061 cgtattaccg ccatgcat
//
