LOCUS       sequence-260011-        5976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for NaVSp1 bacterial sodium 
            channel. Residues 1-224..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5976)
  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 5976)
  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 DeltaCTD pIRES
FEATURES             Location/Qualifiers
     source          1..5976
                     /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(1335..1909,1910..1912,1913..1921)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1502..1519)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1729..1748
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1922..1924,1925..1927,1928..2641)
                     /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(1922..1924,1925..1927,1928..2638)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1967..1988)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2228..2247)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2575..2596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2764..2885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2801..2820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2855..2874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2892..3347)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3029..3050)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3241..3260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3374..3478
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3480..3837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3480..3671
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3503..3523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3688..3823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3872..4666
                     /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             3872..4666
                     /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(3926..3945)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4536..4555
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4854..4873)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4898..4945
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5274..5862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5763..5782
                     /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 tgatgagaat tctgcagtcg acggtaccgc
     1321 gggcccggga tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg
     1381 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     1441 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     1501 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     1561 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     1621 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     1681 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     1741 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     1801 ctggggcctc ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     1861 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     1921 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1981 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2041 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2101 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2161 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2221 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2281 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2341 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2401 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2461 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2521 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2581 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2641 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2701 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2761 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2821 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2881 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2941 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3001 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3061 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3121 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3181 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3241 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3301 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3361 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3421 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3481 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3541 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3601 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3661 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3721 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3781 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3841 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3901 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3961 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4021 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4081 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4141 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4201 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4261 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4321 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4381 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4441 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4501 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4561 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4621 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4681 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4741 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4801 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4861 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4921 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4981 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5041 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5101 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5161 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5221 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5281 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5341 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5401 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5461 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5521 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5581 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5641 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5701 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5761 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5821 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5881 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5941 cctgattctg tggataaccg tattaccgcc atgcat
//
