LOCUS       sequence-468110-        5604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To create cASIC1 channels of defined stoichiometry in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5604)
  AUTHORS   Purohit R, Couch T, Rook ML, MacLean DM
  TITLE     Proline substitutions in the ASIC1 beta11-12 linker slow 
            desensitization.
  JOURNAL   Biophys J. 2024 Oct 15;123(20):3507-3518. doi: 
            10.1016/j.bpj.2024.08.016. Epub 2024 Sep 3.
  PUBMED    39182166
REFERENCE   2  (bases 1 to 5604)
  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.bpj.2024.08.016"; journalName: "Biophys J"; date: 
            "2024-10-15- 15"; volume: "123"; issue: "20"; pages: "3507-3518"
COMMENT     Sequence Label: cASIC1 concatemer - Subunit A
FEATURES             Location/Qualifiers
     source          1..5604
                     /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    2216..2268
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2392..2513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2429..2448)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2483..2502
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2520..2975)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2657..2678)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2869..2888
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3002..3106
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3108..3465
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3108..3299
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3131..3151)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3316..3451
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3500..4294
                     /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             3500..4294
                     /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(3554..3573)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4164..4183
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4482..4501)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4526..4573
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4902..5490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5391..5410
                     /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 gctagcgaag
      601 actagagcca ccatgatgga cctgaaggtg gacgaggagg aggtggacag cgggcagccg
      661 gtgagcatcc aggccttcgc cagcagctcc accctgcacg ggatctcaca catcttctcc
      721 tacgagcggc tgtcgctgaa gcgcgtggtc tgggcgctct gcttcatggg ttcgctggcg
      781 ctgctcgccc tggtctgcac caaccgcatc cagtactact tcctctaccc ccacgtcacc
      841 aagctggacg aggtggccgc caccaggctc accttccctg ctgtcacctt ctgcaacctc
      901 aacgagttcc gcttcagccg ggtgaccaag aacgacctgt accacgccgg ggaactcctc
      961 gccctgctca ataacagata cgagatcccg gacacacaga cagctgatga gaagcagctg
     1021 gaaatcctac aggacaaggc aaacttccga aacttcaagc ccaaaccttt caatatgttg
     1081 gagttttacg accgcgccgg ccacgacatc cgggagatgc tgctctcctg cttcttccgt
     1141 ggggagcagt gcagccctga ggacttcaaa gtggtgttca cacgctatgg gaagtgctac
     1201 acgttcaacg cagggcagga tggcaaaccc cggctcatca ccatgaaggg gggcactgga
     1261 aatggcctgg agatcatgct ggacatccag caggacgagt acctcccagt gtggggggaa
     1321 acagatgaga cctcatttga agccgggatc aaggtgcaga tccacagcca ggatgagcct
     1381 ccactcatcg accagctggg cttcggtgtg gcacccggct tccagacctt cgtgtcctgc
     1441 caggagcagc ggctcatcta cctaccacct ccctggggcg actgtaaggc cacaacgggt
     1501 gactcggagt tctacgacac ttacagcatc actgcctgcc gcatcgactg cgagacccgc
     1561 tacctggtgg agaactgcaa ctgccgcatg gtgcacatgc caggcgatgc cccttattgc
     1621 accccagagc agtacaagga gtgtgcagat ccagccttag atttcctggt ggagaaggac
     1681 aatgagtact gtgtctgtga gatgccctgc aacgtcaccc gctatggcaa agagctctcc
     1741 atggtgaaga tccccagcaa agcctccgcc aagtacctgg caaagaagta caacaagtcg
     1801 gagcagtaca tcggggagaa catcctggtg ctggacatct tctttgaagc cctgaactat
     1861 gagacaatcg agcagaagaa ggcgtatgag gtggctggat tgctgggtga catcggggga
     1921 cagatggggc tgtttatcgg ggccagcatc ctcactgtgc tggagctctt tgactacgcc
     1981 tatgaggtga tcaagcaccg gctgtgccgg cggggcaaat gccgcaagaa ccacaagagg
     2041 aacaacacgg acaagggcgt cgcgctgagc atggatgatg tgaaacgcca caacccctgt
     2101 gagagcctac ggggtcaccc ggctggcatg acgtacgcag cgaacatcct acctcaccac
     2161 ccggcccggg gcacctttga ggacttcacc tgcgcaggta ccgtgaacag tcttcctcga
     2221 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga
     2281 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2341 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2401 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2461 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2521 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2581 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2641 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2701 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2761 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2821 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2881 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2941 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3001 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3061 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3121 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3181 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3241 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3301 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3361 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3421 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3481 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3541 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3601 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3661 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3721 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3781 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3841 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3901 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3961 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4021 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4081 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4141 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4201 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4261 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4321 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4381 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4441 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4501 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4561 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4621 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4681 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4741 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4801 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4861 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4921 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4981 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5041 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5101 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5161 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5221 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5281 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5341 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5401 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5461 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5521 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5581 gataaccgta ttaccgccat gcat
//
