LOCUS       sequence-388133-        7312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control plasmid for the MscL tension biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7312)
  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 7312)
  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)-c-term-cpGFP-ERexp
FEATURES             Location/Qualifiers
     source          1..7312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     383..403
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    513..561
                     /note="synthetic polyadenylation signal"
     CDS             join(827..895,896..1687)
                     /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(1045..1064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1700..1836
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1988..2059
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2064..2110)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2128..2176
                     /note="synthetic polyadenylation signal"
     enhancer        2257..2560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2562..2764
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2715..2735
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3493..3514
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3881..3900)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4108..4229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4119..4138)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4173..4192
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4344..4390
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4420..4556
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4601..4672
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4898..5486)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4978..4997)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5986..7195
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7032..7051
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(7199..7312,1..486)
                     /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(7199..7218)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
       61 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
      121 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
      181 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
      241 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
      301 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
      361 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
      421 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
      481 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
      541 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
      601 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
      661 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
      721 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
      781 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
      841 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      901 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
      961 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     1021 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1081 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     1141 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1201 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     1261 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1321 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1381 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     1441 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1501 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1561 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1621 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1681 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     1741 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     1801 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     1861 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     1921 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     1981 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     2041 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     2101 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     2161 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     2221 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2281 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2341 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2401 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2461 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2521 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     2581 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2641 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2701 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2761 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     2821 ccgccagcgg ccgctgccaa gcttccgagc tctcgaattc atgagcatta ttaaagaatt
     2881 tcgcgaattt gcgatgcgcg ggaacgtggt ggatttggcg gtgagcgtca ttatcggtgc
     2941 ggcattcggg aagattgtct cttcactggt tgccgatatc atcatgcctc ctctgggctt
     3001 attaattggc gggatcgatt ttaaacagtt tgctgtcacg ctacgcgatg cgcaggggga
     3061 tatccctgct gttgtgatgc attacggtgt cttcattcaa aacgtctttg attttctgat
     3121 tgtggccttt gccatcttta tggcgattaa gctaatcaac aaactgaatc ggaaaaaaga
     3181 agaaccagca gccgcacctg caccaactaa agaagaagta ttactgacag aaattcgtga
     3241 tttgctgaaa gagcagaata accgctctgg aagcggagga tccgggaacg tctatatcaa
     3301 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     3361 cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt
     3421 gctgctgccc gacaaccact acctgagcgt gcagtccata ctttcgaaag accccaacga
     3481 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3541 ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt
     3601 caccggggtg gtgcccatcc aggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3661 cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3721 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3781 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3841 gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3901 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3961 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaaca agaaaggcag
     4021 cttttgctat gaaaacgaag tgggatccgc aggcctctgc tagcttgact gactgagata
     4081 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
     4141 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4201 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4261 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     4321 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
     4381 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
     4441 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     4501 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
     4561 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
     4621 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
     4681 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
     4741 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
     4801 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     4861 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4921 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4981 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5041 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5101 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5161 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5221 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5281 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5341 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5401 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5461 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5521 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5581 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5641 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5701 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     5761 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     5821 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     5881 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     5941 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     6001 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     6061 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     6121 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     6181 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     6241 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     6301 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     6361 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     6421 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     6481 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     6541 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     6601 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     6661 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     6721 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     6781 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     6841 gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg
     6901 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     6961 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     7021 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     7081 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     7141 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     7201 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     7261 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg at
//
