LOCUS       sequence-260111-        6156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for NaVAe1 1-264 chimera bacterial 
            sodium channel..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6156)
  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 6156)
  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: NaVAe1 DeltaCC pIRES
FEATURES             Location/Qualifiers
     source          1..6156
                     /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(1515..2089,2090..2092,2093..2101)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1682..1699)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1909..1928
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2102..2104,2105..2107,2108..2821)
                     /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(2102..2104,2105..2107,2108..2818)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2147..2168)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2408..2427)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2755..2776
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2944..3065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2981..3000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3035..3054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3072..3527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3209..3230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3421..3440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3554..3658
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3660..4017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3660..3851
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3683..3703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3868..4003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4052..4846
                     /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             4052..4846
                     /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(4106..4125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4716..4735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5034..5053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5078..5125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5454..6042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5943..5962
                     /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 gctagcatga
      601 gcgagcggca accggatttg gtggggcata aagtacagca cccggaggat gagaccctgc
      661 gtggtcgcct ggcctggttt atcgaccggc cggggacgca gtactttatc gtcggcctga
      721 tcctggtcaa tgcgatcacc ctcgggctga tgacctcgcc ggaggtcacc gcctacctgc
      781 agccctggtt gggctgggtg aacaccttta tcatcgccgc cttcgtggtg gagatctccc
      841 tgcgaatcat cgccgatggg ccccggttcg tgcgcagcgg ttggaatctg ttcgacttct
      901 ccgtagtggc gatctctctg gtgccggact ctggggcctt ctcggtgctg cgggcgctgc
      961 gcatcctgaa ggtgttgcga ctgttctcga tggtcccgcg cctgcgcagg atcgtcgagg
     1021 cgttgctccg cgccatcccg gggatcgcct ggatcgccct gctgctgttg gtgatcttct
     1081 acgtgttcgc ggtgatgggg acgaagctct ttgcccagag cttcccggag tggttcggca
     1141 ccctcggggc cagcatgtac acgctgttcc aggtgatgac cctggagagc tggtccatgg
     1201 gcatcgcccg cccggtgatc gaggcctacc cctgggcctg gatctatttc gtaagcttta
     1261 tcctggtctc ctccttcacg gtgctgaacc tgttcatcgg catcatcatc gagagcatgc
     1321 aatccgccca ctgggaggcg gaggacgcca agcggatcga gcaggagcag cgggcccacg
     1381 acgagcgcta gtagatgctg cagttgatcc gcgatctcag cagcaaggtc gaccggctgg
     1441 aacgtcgctc cggcaagcgg taagagctca agcttcgaat tctgcagtcg acggtaccgc
     1501 gggcccggga tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg
     1561 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     1621 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     1681 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     1741 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     1801 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     1861 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     1921 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     1981 ctggggcctc ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     2041 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     2101 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2161 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2221 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2281 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2341 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2401 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2461 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2521 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2581 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2641 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2701 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2761 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2821 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2881 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2941 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3001 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3061 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3121 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3181 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3241 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3301 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3361 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3421 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3481 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3661 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3901 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3961 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4021 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4081 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4141 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4201 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4261 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4321 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4381 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4441 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4501 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4561 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4621 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4681 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4741 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4801 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4861 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4921 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4981 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5041 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5101 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5161 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5221 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5281 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5341 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5401 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5461 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5521 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5581 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5641 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5701 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5761 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5821 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5881 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5941 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6001 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6061 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6121 cctgattctg tggataaccg tattaccgcc atgcat
//
