LOCUS       sequence-192452-        7101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Electrophysiology in mammalian cells: Mammalian expression of 
            voltage gated sodium channel and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7101)
  AUTHORS   Sula A, Booker J, Ng LC, Naylor CE, DeCaen PG, Wallace BA
  TITLE     The complete structure of an activated open sodium channel.
  JOURNAL   Nat Commun. 2017 Feb 16;8:14205. doi: 10.1038/ncomms14205.
  PUBMED    28205548
REFERENCE   2  (bases 1 to 7101)
  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"; journalName: "Nat 
            Commun."; date: "2017-02-16"; volume: "8:14205. doi"; pages: " 
            10.1038/ncomms14205"
COMMENT     Sequence Label: His-NavMs
FEATURES             Location/Qualifiers
     source          1..7101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      729..738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             747..764
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             774..791
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1670..1687)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1676..1787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1955..3133
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2186..3124
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3081..3101
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        3150..3168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3257..3304
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(3371..3399)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3994..4014
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4037..4414
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MDAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWG
                     REFALRDPAGNCVHFVAEEQD"
     polyA_signal    4544..4665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4581..4600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4635..4654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4714..4730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4714..4730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4727..4749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4738..4754
                     /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)."
     promoter        complement(join(4762..4768,4769..4786,4787..4792))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4807..4828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4945..4962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5116..5704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5196..5215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5875..6666,6667..6735))
                     /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     6498..6517
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6736..6840)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6915..6934)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      601 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      661 gctggctagc gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt
      721 ggaattcagc caccatgggc agcagccatc atcatcatca tcacagcagc ggcctggtgc
      781 cgcgcggcag ccatatgtca cgcaaaatcc gcgatttaat cgaatccaaa cgctttcaaa
      841 acgtcatcac cgccattatt gtgctcaatg gcgctgtgct gggtctgctg accgatacaa
      901 ccctgtcggc ctccagccaa aacctgctgg agcgtgtgga tcaactttgt ctgactatct
      961 ttattgttga aatctccctg aaaatctacg cctatggcgt gcgcggcttt ttccgcagcg
     1021 gctggaatct gtttgatttt gtgattgtgg ccatcgcgct tatgccggcc cagggtagcc
     1081 tgtcggtgct gcgtaccttc cgtatcttcc gcgtcatgcg cctcgtatcg gtcatcccaa
     1141 ccatgcgccg tgtggtgcaa ggcatgctct tggcactgcc gggcgtgggc tcggtagcgg
     1201 cactgttgac ggtggtcttc tatattgcgg ctgtcatggc caccaatctc tacggggcaa
     1261 ccttccctga atggtttggt gatcttagca agagcctgta cacactgttt caggtgatga
     1321 ccttagagtc atggtctatg ggcattgtgc gtccagtgat gaacgttcat ccgaacgcat
     1381 gggttttttt catcccgttc atcatgctca ccacctttac cgtgctcaac ctgtttattg
     1441 gcattattgt agatgcaatg gcaatcacca aggaacagga ggaagaggcc aaaaccggtc
     1501 accatcaaga acctatttct caaactcttc ttcatcttgg tgatcgtctt gatcgtattg
     1561 aaaaacaact tgctcaaaat aatgaacttc ttcaacgtca acaacctcaa aaaaaataag
     1621 gatccgcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1681 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1741 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1801 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1861 gacaatagca gctttgcaaa gatggataaa gttttaaaca gagaggaatc tttgcagcta
     1921 atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
     1981 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
     2041 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     2101 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     2161 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
     2221 ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat
     2281 tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg
     2341 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     2401 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     2461 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     2521 agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     2581 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     2641 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     2701 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     2761 tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     2821 tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc
     2881 cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc
     2941 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     3001 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     3061 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     3121 tcaggtgtcg tgaggaatta gcttggtact aatacgactc actataggga gacccaagct
     3181 ggctaggtaa gctcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3241 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     3301 cgacaaggtg aggaactaaa ccatggcggt agaaaaaatg gctagcaaag gagaagaact
     3361 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
     3421 ttctgtcagt ggagagggtg aaggtgatgc tacatacgga aagcttaccc ttaaatttat
     3481 ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt tctcttatgg
     3541 tgttcaatgc ttttcccgtt atccggatca tatgaaacgg catgactttt tcaagagtgc
     3601 catgcccgaa ggttatgtac aggaacgcac tatatctttc aaagatgacg ggaactacaa
     3661 gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaagg
     3721 tattgatttt aaagaagatg gaaacattct cggacacaaa ctcgagtaca actataactc
     3781 acacaatgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat
     3841 tcgtcacaac attgaagatg gatccgttca actagcagac cattatcaac aaaatactcc
     3901 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgccct
     3961 ttcgaaagat cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc
     4021 tgggattaca catggcatgg atgccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
     4081 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
     4141 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
     4201 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
     4261 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
     4321 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
     4381 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
     4441 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4501 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4561 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4621 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4681 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4741 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4801 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4861 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4921 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4981 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5041 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5101 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5161 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5221 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5281 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5341 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5401 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5461 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5521 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5581 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5641 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5701 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5761 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5821 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5881 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5941 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6001 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6061 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6121 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6181 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6241 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6301 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6361 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6421 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6481 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6541 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6601 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6661 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6721 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6781 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6841 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6901 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     6961 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7021 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     7081 gatgtacggg ccagatatac g
//
