LOCUS       sequence-197873-        7810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the F40-based tension sensor module. The biosensor allows 
            force measurements at 1-6 pN..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7810)
  AUTHORS   Austen K, Ringer P, Mehlich A, Chrostek-Grashoff A, Kluger C, 
            Klingner C, Sabass B, Zent R, Rief M, Grashoff C
  TITLE     Extracellular rigidity sensing by talin isoform-specific mechanical 
            linkages.
  JOURNAL   Nat Cell Biol. 2015 Dec;17(12):1597-606. doi: 10.1038/ncb3268. Epub 
            2015 Nov 2.
  PUBMED    26523364
REFERENCE   2  (bases 1 to 7810)
  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/ncb3268"; 
            journalName: "Nat Cell Biol"; date: "2015-12"; volume: "17"; issue: 
            "12"; pages: "1597-606"
COMMENT     Sequence Label: F40 tension sensor
FEATURES             Location/Qualifiers
     source          1..7810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1565..2164
                     /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(1565..1584)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2061..2081
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2339..2642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2643..2846
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2797..2817
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(3239..3258)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             3749..4456
                     /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(3894..3912)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4162..4181
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4469..4486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     LTR             4549..5142
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5245..5267)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5395..5428
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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)."
     primer_bind     5541..5560
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5641..5658)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5812..6400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5892..5911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6571..7362,7363..7431))
                     /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     7194..7213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7432..7536)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7604..7622
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aatgatcccc cgggctgcag gtcggccgcc
     1501 acgaccggtg ccgccaccat cccctgaccc acgcccctga cccctcacaa ggagacgacc
     1561 ttccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     1621 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     1681 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     1741 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     1801 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     1861 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     1921 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     1981 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     2041 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     2101 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     2161 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc
     2221 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac
     2281 tctagtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2341 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2401 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2461 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2521 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2581 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2641 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     2701 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2761 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     2821 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     2881 ggactcagat ctgggcccat aaagcttata cgaattcacc atggcttgta aaatggtgag
     2941 caaaggcgaa gagctgttca ccggcgtggt gcccatcctg gtggagctgg acggcgacgt
     3001 gaacggccac aagttcagcg tgagcggcga gggcgagggc gacgccacct acggcaagct
     3061 gaccctgaag ctgctgtgca ccaccggcaa gctgcccgtg ccctggccca ccctggtgac
     3121 caccctgggc tacggcgtgc agtgcttcgc ccggtacccc gaccacatga agcagcacga
     3181 cttcttcaag agcgccatgc ccgagggcta cgtgcaggag cggaccatct tcttcaagga
     3241 cgacggcaac tacaagaccc gggccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3301 gatcgagctg aagggcatcg acttcaagga ggacggcaac atcctgggcc acaagctgga
     3361 gtacaactac aacagccaca acgtgtacat caccgccgac aagcagaaga acggcatcaa
     3421 ggccaacttc aagatccggc acaacatcga ggacggcggc gtgcagctgg ccgaccacta
     3481 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3541 ctaccagagc gccctgttca aggaccccaa cgagaagcgg gaccacatgg tgctgctgga
     3601 gttcctgacc gccgccggat ccgggccagg tggagcaggg ccaggtggtg cagggccagg
     3661 tggtgcaggg ccaggtggtg cagggcccgg tggtgcaggt ccaggtggtg caggtccagg
     3721 tggtgctggt ccaggtggtg ctggatccat ggtgagcaag ggcgaggagg ataacatggc
     3781 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     3841 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3901 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     3961 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     4021 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     4081 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     4141 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     4201 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     4261 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     4321 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     4381 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     4441 ggacgagctg tacaagaaat gtctcgagca ccaccaccac caccactgag cggccgccta
     4501 tacatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     4561 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     4621 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     4681 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4741 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4801 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     4861 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     4921 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     4981 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     5041 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     5101 tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat
     5161 cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg gctgcctcgc
     5221 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5281 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5341 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt actgataact
     5401 tcgtataatg tatgctatac gaagttatta ggtctgaaga ggagtttacg tccagccaag
     5461 ctagggccgc gatccggaac ccttaatata acttcgtata atgtatgcta tacgaagtta
     5521 tcagtactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5581 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     5641 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5701 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5761 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5821 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5881 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5941 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6001 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6061 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6241 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6361 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6421 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6481 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6541 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6601 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6661 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6721 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6781 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6841 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt
     6901 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6961 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7021 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7081 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7141 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     7201 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7261 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7321 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7381 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7441 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7501 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7561 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7621 gccctttcgt cttcaagaat tgctagcaat tgctagcaat tgctagcaat tcataccaga
     7681 tcaccgaaaa ctgtcctcca aatgtgtccc cctcacactc ccaaattcgc gggcttctgc
     7741 ctcttagacc actctaccct attccccaca ctcaccggag ccaaagccgc ggcccttccg
     7801 tttctttgct
//
