LOCUS       sequence-450556-        7119 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses a mutant of the Arcobacter butzleri voltage-gated sodium 
            channel fused to an MBP tag in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7119)
  AUTHORS   Wisedchaisri G, Tonggu L, McCord E, Gamal El-Din TM, Wang L, Zheng 
            N, Catterall WA
  TITLE     Resting-State Structure and Gating Mechanism of a Voltage-Gated 
            Sodium Channel.
  JOURNAL   Cell. 2019 Aug 8;178(4):993-1003.e12. doi: 
            10.1016/j.cell.2019.06.031. Epub 2019 Jul 25.
  PUBMED    31353218
REFERENCE   2  (bases 1 to 7119)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2019.06.031"; journalName: "Cell"; date: "2019-08-8-
            8"; volume: "178"; issue: "4"; pages: "993-1003.e12"
COMMENT     Sequence Label: pFB-Dual-MBP-NavAb (KAV/G94C/Q150C)
FEATURES             Location/Qualifiers
     source          1..7119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(415..420,421..437,438..443))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    963..1011
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1036..1055
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(1147..1256)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1314..1405
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1330..1347
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1470..2570
                     /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     2544..2567
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     polyA_signal    3513..3647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3550..3569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3604..3623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3682..3847
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4031..4485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4118..4137)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4327..4348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4511..4615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4616..4684,4685..5476)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4834..4853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5647..6235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6136..6155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6389..6406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6540..6764
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(6831..7119,1..245))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc
       61 ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca
      121 gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt
      181 gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat
      241 aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac
      301 tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg
      361 cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta
      421 cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca cggtgtgcgt
      481 cacccggcaa ccttgggcag cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag
      541 cgcaaggttt cggtctccac gcatcgtcag gcattggcgg ccttgctgtt cttctacggc
      601 aaggtgctgt gcacggatct gcccttgctt caggagatcg gtagacctcg gccgtcgcgg
      661 cgcttgccgg tggtgctgac cccggatgaa gtggttcgca tcctcggttt tctggaaggc
      721 gagcatcgtt tgttcgccca ggactctagc tatagttcta gtggttggct acgtatcgcg
      781 aggcgccccc gtagtggcta tggcagggct tgccgccccg acgttggctg cgagccctgg
      841 gccttcaccc gaacttgggg gttggggtgg ggaaaaggaa gaaacgcggg cgtattggtc
      901 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
      961 atgaacaaac gacccaacac ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     1021 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccat ctcccggtac
     1081 cgcatgctat gcatcagctg ctagcaccat ggctcgagat cccgggtgat caagtcttcg
     1141 tcgagtgatt gtaaataaaa tgtaatttac agtatagtat tttaattaat atacaaatga
     1201 tttgataata attcttattt aactataata tattgtgttg ggttgaatta aaggtccgta
     1261 tacgtttaaa catcgatcac gtgactagtg gcgcgcctcc ggaatattaa tagatcatgg
     1321 agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt ttcgtaacag
     1381 ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca ccatcgggcg
     1441 cggatcccgg tccgaagcgc gcggaattca tgaaaatcga agaaggtaaa ctggtaatct
     1501 ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag
     1561 ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg
     1621 ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct
     1681 acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt
     1741 atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg
     1801 ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg
     1861 aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca
     1921 acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca
     1981 agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag
     2041 cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt
     2101 actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt
     2161 gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct
     2221 tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca
     2281 gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc
     2341 tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag
     2401 agttggcgaa agatccacgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca
     2461 tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg
     2521 ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aatgctatgt
     2581 atcttgcaat tacaaatatt gtagaaagta gctttttcac aaagtttata atttatttaa
     2641 ttgtattaaa tggaattaca atgggacttg agacttcaaa aacttttatg caaagttttg
     2701 gagtttatac aacgctattt aaacaaattg ttattactat ttttactata gagattattt
     2761 taagaattta tgttcataga atctcttttt tcaaagaccc atggagtcta tttgattttt
     2821 tcgttgttgc tatctcttta gttcctacaa gttcatgctt tgaaatatta agagttttaa
     2881 gagttcttag actatttagg gcagttactg ctgttcctca agtgagaaaa atagtttcag
     2941 cacttattag tgtgattcca ggaatgttat ctgtaattgc tttaatgaca ctgttttttt
     3001 atatttttgc aattatggca acatgccttt ttggagaaag atttcctgaa tggtttggaa
     3061 cattaggtga atctttttat acactatttc aagttatgac tttagaatct tggtcaatgg
     3121 gtattgtcag acctctgatg gaagtttatc cttatgcttg ggttttcttt atacctttta
     3181 tatttgtagt aacttttgta atgataaatt tagtggttgc tattatagtt gatgcaatgg
     3241 ctattttaaa tcaaaaagag gaacaacata taattgatga agttcaatct catgaagata
     3301 atataaataa tgagataata aaattaagag aagagattgt agaacttaaa gaattaataa
     3361 aaactagttt aaaaaactaa taaaagcttg tcgagaagta ctagaggatc ataatcagcc
     3421 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3481 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg
     3661 agacgcgtcc taggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca
     3721 tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct
     3781 tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc
     3841 gcccacagcg gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca
     3901 tgtgacaaac cgtcatcttc ggctactttt tctctgtcac agaatgaaaa tttttctgtc
     3961 atctcttcgt tattaatgtt tgtaattgac tgaatatcaa cgcttatttg cagcctgaat
     4021 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4081 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4141 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4201 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4261 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4321 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4381 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4441 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4501 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4561 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4621 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4681 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4741 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4801 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4861 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4921 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4981 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5041 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5101 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5161 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5221 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5281 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5341 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5401 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5461 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5521 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5581 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5641 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5701 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5761 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5821 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5881 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5941 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6001 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6061 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6121 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6181 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6241 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6301 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6361 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6421 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagaccagcc
     6481 gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg gatgccctgc gtaagcgggt
     6541 gtgggcggac aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc
     6601 ttaaactaga cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata
     6661 ctggactttt gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg
     6721 tattaaagag gggcgtggcc aagggcatgg taaagactat attcgcggcg ttgtgacaat
     6781 ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg
     6841 gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga
     6901 gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg
     6961 ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg
     7021 ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggcaga
     7081 acgtaagccg cgagagcgcc aacaaccgct tcttggtcg
//
