LOCUS       sequence-384238-        7282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter/biosensor for membrane tension.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7282)
  AUTHORS   Hsu YY, Resto Irizarry AM, Fu J, Liu AP
  TITLE     Mechanosensitive Channel-Based Optical Membrane Tension Reporter.
  JOURNAL   ACS Sens. 2023 Jan 27;8(1):12-18. doi: 10.1021/acssensors.2c01921. 
            Epub 2023 Jan 6.
  PUBMED    36608338
REFERENCE   2  (bases 1 to 7282)
  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: 
            "10.1021/acssensors.2c01921"; journalName: "ACS Sens"; date: 
            "2023-01-27- 27"; volume: "8"; issue: "1"; pages: "12-18"
COMMENT     Sequence Label: MscL(G22S)61-cpGFP-ERexp
FEATURES             Location/Qualifiers
     source          1..7282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     376..396
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    506..554
                     /note="synthetic polyadenylation signal"
     CDS             join(820..888,889..1680)
                     /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(1038..1057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1693..1829
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1981..2052
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2057..2103)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2121..2169
                     /note="synthetic polyadenylation signal"
     enhancer        2250..2553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2555..2757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2708..2728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3240..3261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3628..3647)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4071..4192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4082..4101)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4136..4155
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4307..4353
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4383..4519
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4564..4635
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4861..5449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4941..4960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5949..7158
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6995..7014
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(7162..7282,1..479)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7162..7181)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
       61 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
      121 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
      181 cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg cccaaggagc
      241 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
      301 gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg
      361 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
      421 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg
      481 ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt
      541 tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg
      601 cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac
      661 ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg
      721 tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca
      781 aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt
      841 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      901 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg
      961 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg
     1021 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1081 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca
     1141 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1201 agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc
     1261 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1321 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc
     1381 ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac
     1441 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1501 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1561 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1621 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1681 ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     1741 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     1801 tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct
     1861 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     1921 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     1981 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     2041 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     2101 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     2161 gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca
     2221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2521 tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat
     2581 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2641 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2701 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     2761 ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag
     2821 cggccgctgc caagcttccg agctctcgaa ttcatgagca ttattaaaga atttcgcgaa
     2881 tttgcgatgc gcgggaacgt ggtggatttg gcggtgagcg tcattatcgg tgcggcattc
     2941 gggaagattg tctcttcact ggttgccgat atcatcatgc ctcctctggg cttattaatt
     3001 ggcgggatcg attttaaaca gtttgctgtc acgaacgtct atatcaaggc cgacaagcag
     3061 aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3121 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3181 aaccactacc tgagcgtgca gtccatactt tcgaaagacc ccaacgagaa gcgcgatcac
     3241 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3301 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
     3361 cccatccagg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3421 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3481 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     3541 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3601 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3661 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3721 gacggcaaca tcctggggca caagctggag tacaacctac gcgatgcgca gggggatatc
     3781 cctgctgttg tgatgcatta cggtgtcttc attcaaaacg tctttgattt tctgattgtg
     3841 gcctttgcca tctttatggc gattaagcta atcaacaaac tgaatcggaa aaaagaagaa
     3901 ccagcagccg cacctgcacc aactaaagaa gaagtattac tgacagaaat tcgtgatttg
     3961 ctgaaagagc agaataaccg ctctttttgc tatgaaaatg aagttggatc cgcaggcctc
     4021 tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
     4081 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4141 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4201 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4261 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     4321 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     4381 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     4441 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     4501 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     4561 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     4621 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     4681 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     4741 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     4801 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4861 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4981 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5041 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5101 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5161 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5221 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5281 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5341 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5461 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5521 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5581 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5641 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     5701 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     5761 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     5821 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     5881 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     5941 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     6001 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     6061 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     6121 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     6181 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     6241 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     6301 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     6361 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     6421 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     6481 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     6541 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     6601 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     6661 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     6721 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     6781 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     6841 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     6901 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     6961 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     7021 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     7081 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     7141 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     7201 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     7261 cgccacaccg tcgatccgga cc
//
