LOCUS       sequence-197870-        7936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the FL-based tension sensor module. The biosensor allows 
            force measurements at 3-5 pN..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7936)
  AUTHORS   Ringer P, Weissl A, Cost AL, Freikamp A, Sabass B, Mehlich A, 
            Tramier M, Rief M, Grashoff C
  TITLE     Multiplexing molecular tension sensors reveals piconewton force 
            gradient across talin-1.
  JOURNAL   Nat Methods. 2017 Nov;14(11):1090-1096. doi: 10.1038/nmeth.4431. 
            Epub 2017 Sep 18.
  PUBMED    28945706
REFERENCE   2  (bases 1 to 7936)
  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: 
            "10.1038/nmeth.4431"; journalName: "Nat Methods"; date: "2017-11"; 
            volume: "14"; issue: "11"; pages: "1090-1096"
COMMENT     Sequence Label: FL tension sensor module
FEATURES             Location/Qualifiers
     source          1..7936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             381..556
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    619..818
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1019..1435
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1365..1387
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1403..1419
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1532..2131
                     /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(1532..1551)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2028..2048
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2306..2609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2610..2813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2764..2784
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(3206..3225)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             3590..3835
                     /codon_start=1
                     /product="mechanically labile synthetic ferredoxin-like 
                     domain (Fang et al., 2013)"
                     /label=FL
                     /note="can serve as an intracellular tension sensor (Ringer
                     et al., 2017)"
                     /translation="MGEFDIRFRTDDDEQFEKVLKEMNRRARKDAGTVTYTRDGNDFEI
                     RITGISEQNRKELAKEVERLAKEQNITVTYTERGSLE"
     CDS             3842..4549
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3987..4005)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4255..4274
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4562..4579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     LTR             4642..5235
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5338..5360)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5488..5521
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5581..5614
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5634..5653
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5734..5751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5905..6493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5985..6004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6664..7455,7456..7524))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7287..7306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7525..7629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7697..7715
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttaagtaac gccactttgc aaggcatgga aaaatacata actgagaata gaaaagttca
       61 gatcaaggtc aggaacaaag aaacagctga ataccaaaca ggatatctgt ggtaagcggt
      121 tcctgccccg gctcagggcc aagaacagat gagacagctg agtgatgggc caaacaggat
      181 atctgtggta agcagttcct gccccggctc ggggccaaga acagatggtc cccagatgcg
      241 gtccagccct cagcagtttc tagtgaatca tcagatgttt ccagggtgcc ccaaggacct
      301 gaaaatgacc ctgtacctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      361 gcgcttccgc tctccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      421 ttccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca
      481 tccgaatcgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccacg
      541 acgggggtct ttcatttggg ggctcgtccg ggatttggag acccctgccc agggaccacc
      601 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      661 gtctatgttt gatgttatgc gcctgcgtct gtactagtta gctaactagc tctgtatctg
      721 gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc
      781 cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc
      841 gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
      901 gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg
      961 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg
     1021 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     1081 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     1141 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1201 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1261 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1321 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1381 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat
     1441 gatcccccgg gctgcaggtc ggccgccacg accggtgccg ccaccatccc ctgacccacg
     1501 cccctgaccc ctcacaagga gacgaccttc catgaccgag tacaagccca cggtgcgcct
     1561 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     1621 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     1681 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     1741 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     1801 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     1861 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     1921 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     1981 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     2041 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     2101 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     2161 cgaaaggagc gcacgacccc atggctccga ccgaagccac ccggggcggc cccgccgacc
     2221 ccgcacccgc ccccgaggcc caccgactct agtaatagta atcaattacg gggtcattag
     2281 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2341 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2401 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2461 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2521 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2581 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2641 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2701 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2761 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     2821 tgaaccgtca gatccgctag cgctaccgga ctcagatctg ggcccataaa gcttatacga
     2881 attcaccatg gcttgtaaaa tggtgagcaa aggcgaagag ctgttcaccg gcgtggtgcc
     2941 catcctggtg gagctggacg gcgacgtgaa cggccacaag ttcagcgtga gcggcgaggg
     3001 cgagggcgac gccacctacg gcaagctgac cctgaagctg ctgtgcacca ccggcaagct
     3061 gcccgtgccc tggcccaccc tggtgaccac cctgggctac ggcgtgcagt gcttcgcccg
     3121 gtaccccgac cacatgaagc agcacgactt cttcaagagc gccatgcccg agggctacgt
     3181 gcaggagcgg accatcttct tcaaggacga cggcaactac aagacccggg ccgaggtgaa
     3241 gttcgagggc gacaccctgg tgaaccggat cgagctgaag ggcatcgact tcaaggagga
     3301 cggcaacatc ctgggccaca agctggagta caactacaac agccacaacg tgtacatcac
     3361 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccggcaca acatcgagga
     3421 cggcggcgtg cagctggccg accactacca gcagaacacc cccatcggcg acggccccgt
     3481 gctgctgccc gacaaccact acctgagcta ccagagcgcc ctgttcaagg accccaacga
     3541 gaagcgggac cacatggtgc tgctggagtt cctgaccgcc gccggatcta tgggcgagtt
     3601 tgacatccgg tttcggactg atgacgacga acagttcgag aaagtgctga aggagatgaa
     3661 tcgtcgagcc agaaaggatg ctggaactgt gacctacaca agggatggga atgacttcga
     3721 gattcgcatt accggcataa gcgagcaaaa ccgcaaagaa ctggccaaag aggttgaaag
     3781 gcttgcaaag gaacagaaca tcacagtcac gtataccgag agaggttccc tcgaaggatc
     3841 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     3901 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     3961 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     4021 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4081 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     4141 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     4201 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     4261 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     4321 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4381 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     4441 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     4501 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga aatgtctcga
     4561 gcaccaccac caccaccact gagcggccgc ctatacatcg ataaaataaa agattttatt
     4621 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     4681 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc
     4741 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     4801 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     4861 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     4921 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     4981 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     5041 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     5101 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     5161 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5221 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
     5281 acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     5341 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5401 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     5461 gtcacgtagc gatagcggag tgtactgata acttcgtata atgtatgcta tacgaagtta
     5521 ttaggtctga agaggagttt acgtccagcc aagctagggc cgcgatccgg aacccttaat
     5581 ataacttcgt ataatgtatg ctatacgaag ttatcagtac tggcttaact atgcggcatc
     5641 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     5701 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5761 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5821 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5881 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5941 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6001 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6061 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6421 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6481 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6541 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6601 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6661 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6721 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6781 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6841 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6901 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6961 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7021 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7081 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7141 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7201 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7261 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     7321 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7381 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7441 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7501 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7621 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7681 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattgctagc
     7741 aattgctagc aattgctagc aattcatacc agatcaccga aaactgtcct ccaaatgtgt
     7801 ccccctcaca ctcccaaatt cgcgggcttc tgcctcttag accactctac cctattcccc
     7861 acactcaccg gagccaaagc cgcggccctt ccgtttcttt gcttttgaaa gaccccaccc
     7921 gtaggtggca agctag
//
