LOCUS       sequence-260010-        7309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for NaVSp1 bacterial sodium channel. It 
            expresses for HIStag-MBP-3C-channel protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7309)
  AUTHORS   Shaya D, Kreir M, Robbins RA, Wong S, Hammon J, Bruggemann A, Minor 
            DL Jr
  TITLE     Voltage-gated sodium channel (NaV) protein dissection creates a set 
            of functional pore-only proteins.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Jul 26;108(30):12313-8. doi: 
            10.1073/pnas.1106811108. Epub 2011 Jul 11.
  PUBMED    21746903
REFERENCE   2  (bases 1 to 7309)
  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: 
            "10.1073/pnas.1106811108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-07-26- 26"; volume: "108"; issue: "30"; pages: "12313-8"
COMMENT     Sequence Label: HM3C NaVSp1 pET24b
FEATURES             Location/Qualifiers
     source          1..7309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(203..220)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1013..1036)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(1103..2203)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(1106..1129)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             complement(2213..2230)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2250..2272
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2287..2311
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(2311..2330)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2312..2330)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2414..2433)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2639..2716
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2717..3799
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2736..2755)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4608..4799
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4793..4815)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4901..5040
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4956..4975
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5055..5072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5226..5814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5306..5325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5925..5944)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5936..6751
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5936..6751
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6009..6028)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6843..7298)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6980..7001)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7192..7211
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttaacag catggaccac
      181 agcagtcgac cgagcggccg ccgctaccac ggggaaccaa cccttattac tttttggttt
      241 caccaagcgc attcagccgc tggtcgatct gttccaggcg ggccagaacc tcgtcccgat
      301 aggcatcggt ccgctcgcca tcctcggcgt gatgagcgtc ctgcatcgag ttcacgatca
      361 gaccgaccag caggttaacc accgcaaatg tggtgatcat gatgaagggg acaaagaaca
      421 gccaggcata gggatagacc tgcatcaccg ggcgaacgat gcccatcgac cagctttcca
      481 gcgtcatgat ctggaacagc gtataggccg acagacccag atcgccgaac cagtcgggaa
      541 aggaggcggc gaacagcttg gtcgcgatca ccgcgccgat gtagaagatg atggtcatca
      601 acagaaacac gctggccatg ccgggcagcg cagtgatgaa gccttcgacc acgcggcgca
      661 agcgcggcgc gaccgagatc acgcgaagca cccgcaggat gcgcagcgcc cgcagcaccg
      721 acagaccccc ggcggcgggg atcagggcga tcccgacgat cacgaaatcg aacaggttcc
      781 agccgttcag gaaaaagcgc tgccgctgcg ccgtcagttt cagcgcgatc tcgacgatga
      841 agatgcctag gcagatacgg tcagcggcca ggatcagccc gccccaccgc tccatcagat
      901 agggcgacgt ctccatcccc agcacgacgg cgttcagcat gatcacggcg gtaatgaacc
      961 ggccgaaacc gggccggtcg ataaaggcct gcattctttg ggaggggctg ctcggtcctt
     1021 gaaacaaaac ttccaaagat ccgccatcga tggagaattc gttgttgtta ttgttattgt
     1081 tgttgttgtt cgagctcgaa ttagtctgcg cgtctttcag ggcttcatcg acagtctgac
     1141 gaccgctggc ggcgttgatc accgcagtac gcacggcata ccagaaagcg gacatctgcg
     1201 ggatgttcgg catgatttca cctttctggg cgttttccat agtggcggca atacgtggat
     1261 ctttcgccaa ctcttcctcg taagacttca gcgctacggc acccagcggt ttgtctttat
     1321 taaccgcttc cagaccttca tcagtcagca gatagttttc gaggaactct tttgccagct
     1381 ctttgttcgg actggcggcg ttaatacctg cgctcagcac gccaacgaac ggtttggatg
     1441 gttgaccctt gaaggtcggc agtaccgtta caccataatt cactttgctg gtgtcgatgt
     1501 tggaccatgc ccacgggccg ttgatggtca tcgctgtttc gcctttatta aaggcagctt
     1561 ctgcgatgga gtaatcggtg tctgcattca tgtgtttgtt tttaatcagg tcaaccagga
     1621 aggtcagacc cgctttcgcg ccagcgttat ccacgcccac gtctttaatg tcgtacttgc
     1681 cgttttcata cttgaacgca taacccccgt cagcagcaat cagcggccag gtgaagtacg
     1741 gttcttgcag gttgaacatc agcgcgctct tacctttcgc tttcagttct ttatccagcg
     1801 ccgggatctc ttcccaggtt tttggcgggt tcggcagcag atctttgtta taaatcagcg
     1861 ataacgcttc aacagcgatc gggtaagcaa tcagcttgcc gttgtaacgt acggcatccc
     1921 aggtaaacgg atacagcttg tcctggaacg ctttgtccgg ggtgatttca gccaacaggc
     1981 cagattgagc gtagccacca aagcggtcgt gtgcccagaa gataatgtca gggccatcgc
     2041 cagttgccgc aacctgtggg aatttctctt ccagtttatc cggatgctca acggtgactt
     2101 taattccggt atctttctcg aatttcttac cgacttcagc gagaccgtta tagcctttat
     2161 cgccgttaat ccagattacc agtttacctt cttcgatttt catagaagaa ccatggtgat
     2221 ggtgatggtg agaagaaccc atatgtatat ctccttctta aagttaaaca aaattatttc
     2281 tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg
     2341 atcgagatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     2401 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     2461 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     2521 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     2581 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga
     2641 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     2701 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     2761 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     2821 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     2881 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     2941 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     3001 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     3061 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     3121 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     3181 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     3241 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     3301 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     3361 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     3421 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     3481 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     3541 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     3601 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     3661 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     3721 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3781 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     3841 cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     3901 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     3961 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     4021 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     4081 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc
     4141 cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc
     4201 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca
     4261 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc
     4321 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct
     4381 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     4441 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     4501 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     4561 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg
     4621 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga
     4681 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg
     4741 ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4801 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4861 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     4921 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     4981 agtgcaccat tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5041 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5101 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5161 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5221 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5281 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5341 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5401 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5461 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5521 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5581 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5641 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5701 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5761 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5821 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5881 ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     5941 ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca tggatgctga
     6001 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     6061 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     6121 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     6181 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     6241 tgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     6301 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     6361 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     6421 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     6481 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     6541 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     6601 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     6661 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     6721 gtttcatttg atgctcgatg agtttttcta agaattaatt catgagcgga tacatatttg
     6781 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6841 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     6901 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     6961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     7021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     7081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     7141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     7201 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     7261 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
//
