LOCUS       sequence-207549-        6497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eMscL G22S is optimized for mammalian rodent neuronal expression to 
            the plasma membrane through a neuron-specific promoter and a 
            voltage-gated channel targeting motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  AUTHORS   Soloperto A, Boccaccio A, Contestabile A, Moroni M, Hallinan GI, 
            Palazzolo G, Chad J, Deinhardt K, Carugo D, Difato F
  TITLE     Mechano-sensitization of mammalian neuronal networks through 
            expression of the bacterial mechanosensitive MscL channel.
  JOURNAL   J Cell Sci. 2018 Jan 29. pii: jcs.210393. doi: 10.1242/jcs.210393.
  PUBMED    29361543
REFERENCE   2  (bases 1 to 6497)
  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: "J Cell Sci.
            2018 Jan 29. pii: jcs.210393. doi: 10.1242/jcs.210393."
COMMENT     Sequence Label: eMscL G22S
FEATURES             Location/Qualifiers
     source          1..6497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     655..679
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      693..702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1116..1133
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      1146..1155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1152..2579
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    2616..3204
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2669..2689)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3087..3098)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3244..3720
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3443..3462)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3760..3900
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3760..3889
                     /label=AAV2 ITR
     rep_origin      3975..4430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4062..4081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4272..4293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4447..4466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4566..4588
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4626..4644)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4712..4816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4817..4885,4886..5677)
                     /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(5035..5054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5848..6436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6337..6356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccaccgg tagatctgct agcgagctcg
      661 tttagtgaac cgtcagatcg cctggagaat tcgccaccat ggcgagcatt attaaagaat
      721 ttcgcgaatt tgcgatgcgc gggaacgtgg tggatttggc ggtgagtgtc attatcggtg
      781 cggcattcgg gaagattgtc tcttcactgg ttgccgatat catcatgcct cctctgggct
      841 tattaattgg cgggatcgat tttaaacagt ttgctgtcac gctacgcgat gcgcaggggg
      901 atatccctgc tgttgtgatg cattacggtg tcttcattca aaacgtcttt gattttctga
      961 ttgtggcctt tgccatcttt atggcgatta agctaatcaa caaactgaat cggaaaaaag
     1021 aagaaccagc agccgcacct gcaccaacta aagaagaagt attactgaca gaaattcgtg
     1081 atttgctgaa agagcagaat aaccgctctg ctagcctggt tccgcgtgga tctgtcgaca
     1141 gatccgccac catggtgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg
     1201 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1261 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     1321 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
     1381 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
     1441 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg
     1501 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
     1561 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
     1621 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccgc tacctggtgg
     1681 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
     1741 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     1801 gctccgaggg ccgccaccac ctgttcctgg ggcatggcac cggcagcacc ggcagcggca
     1861 gctccggcac cgcctcctcc gaggacaaca acatggccgt catcaaagag ttcatgcgct
     1921 tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     1981 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     2041 tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga
     2101 agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg
     2161 agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc
     2221 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     2281 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     2341 acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc
     2401 tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact
     2461 acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2521 acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag ctgtacaagt
     2581 tctgctacga gaacgaggtg taaaagctta tcgataatca acctctggat tacaaaattt
     2641 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2701 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2761 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2821 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2881 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2941 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3001 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     3061 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3121 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3181 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgattta attaatgatc
     3241 tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     3301 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     3361 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     3421 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     3481 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     3541 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     3601 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     3661 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     3721 ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3781 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3841 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3901 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3961 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4021 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4081 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4141 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4201 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4261 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4321 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4381 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4441 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4501 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4561 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4621 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4681 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4801 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4861 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4921 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4981 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5041 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5101 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5161 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5221 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5281 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5341 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5401 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5461 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5521 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5581 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5641 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5701 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5761 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5821 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5881 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5941 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6001 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6061 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6121 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6181 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6241 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6301 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6361 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6421 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6481 gccttttgct cacatgt
//
