LOCUS       sequence-260097-        6081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for (NaVSp1 1-220 NaVBh1 236-274) 
            chimera bacterial sodium channel..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6081)
  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 6081)
  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 Bh1 CTD pIRES
FEATURES             Location/Qualifiers
     source          1..6081
                     /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(1440..2014,2015..2017,2018..2026)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1607..1624)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1834..1853
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2027..2029,2030..2032,2033..2746)
                     /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(2027..2029,2030..2032,2033..2743)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2072..2093)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2333..2352)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2680..2701
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2869..2990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2906..2925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2960..2979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2997..3452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3134..3155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3346..3365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3479..3583
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3585..3942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3585..3776
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3608..3628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3793..3928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3977..4771
                     /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             3977..4771
                     /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(4031..4050)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4641..4660
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4959..4978)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5003..5050
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5379..5967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5868..5887
                     /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 actcgatgga aaaagcagag ttaacggaca atgaggaaga tggtgaagcc
     1321 gatgggttaa aacaagaaat ctcagcctta agaaaagacg tagccgagct aaaaagcttg
     1381 cttaaacaat cgaaataata agaattctgc agtcgacggt accgcgggcc cgggatccgc
     1441 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1501 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     1561 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     1621 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     1681 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     1741 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     1801 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     1861 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     1921 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     1981 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     2041 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2101 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2161 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2221 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2281 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2341 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2401 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2461 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2521 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2581 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2641 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2701 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     2761 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2821 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3001 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3061 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3121 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3181 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3241 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3301 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3361 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3421 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3601 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3661 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3721 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3781 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3841 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3901 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3961 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4021 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4081 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4141 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4201 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4261 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4321 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4381 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4441 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4501 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4561 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4621 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4681 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4741 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4801 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4861 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4921 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4981 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5041 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5101 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5161 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5221 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5281 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5341 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5401 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5461 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5521 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5581 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5641 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5701 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5761 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5821 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5881 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5941 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6001 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6061 aaccgtatta ccgccatgca t
//
