LOCUS       sequence-430713-        7266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Actin Binding Domain Tension Sensor (ABDTS), composed of 
            Ftractin-Linker-TSMod-Linker-Ftractin. TSMod is the tension sensor 
            module containing mTFP1 and mVenus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7266)
  AUTHORS   Shoyer TC, Collins KL, Ham TR, Blanchard AT, Malavade JN, Johns BA, 
            West JL, Hoffman BD
  TITLE     Detection of fluorescent protein mechanical switching in cellulo.
  JOURNAL   Cell Rep Methods. 2024 Jul 15;4(7):100815. doi: 
            10.1016/j.crmeth.2024.100815. Epub 2024 Jul 9.
  PUBMED    38986612
REFERENCE   2  (bases 1 to 7266)
  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"; journalName: "Cell Rep 
            Methods."; date: "2024-07-15"; volume: "4(7):100815. doi"; pages: " 
            10.1016/j.crmeth.2024.100815. Epub 2024 Jul 9"
COMMENT     Sequence Label: Actin Binding Domain Tension Sensor
FEATURES             Location/Qualifiers
     source          1..7266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1109..1816
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             join(1937..1939,1940..1942,1943..2653)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1982..2003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2243..2262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2590..2611
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2681..2690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2857..2874)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2863..3087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3133..3561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3220..3239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3430..3451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3570..3590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3575..3904
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3755..3890
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3817..3836
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3971..4765
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4025..4044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4635..4654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4939..5060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4976..4995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5030..5049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5109..5125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5109..5125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5122..5144)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5133..5149
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5157..5163,5164..5181,5182..5187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5202..5223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5340..5357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5511..6099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5591..5610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6270..7061,7062..7130))
                     /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     6893..6912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7131..7235)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggatggcgcg
      961 accacggggc gcggggccct gcagccccgg gttggagcgg gctccgcgcc ggagcgtcgg
     1021 ggagctgcgc ctgctcttcg aagcgcgctg cgccgcagtc gccgccgcag cggccgcagg
     1081 ggggctagcg ctaccggtcg ccaccatgat ggtgagcaag ggcgaggaga ccacaatggg
     1141 cgtaatcaag cccgacatga agatcaagct gaagatggag ggcaacgtga atggccacgc
     1201 cttcgtgatc gagggcgagg gcgagggcaa gccctacgac ggcaccaaca ccatcaacct
     1261 ggaggtgaag gagggagccc ccctgccctt ctcctacgac attctgacca ccgcgttcgc
     1321 ctacggcaac agggccttca ccaagtaccc cgacgacatc cccaactact tcaagcagtc
     1381 cttccccgag ggctactctt gggagcgcac catgaccttc gaggacaagg gcatcgtgaa
     1441 ggtgaagtcc gacatctcca tggaggagga ctccttcatc tacgagatac acctcaaggg
     1501 cgagaacttc ccccccaacg gccccgtgat gcagaagaag accaccggct gggacgcctc
     1561 caccgagagg atgtacgtgc gcgacggcgt gctgaagggc gacgtcaagc acaagctgct
     1621 gctggagggc ggcggccacc accgcgttga cttcaagacc atctacaggg ccaagaaggc
     1681 ggtgaagctg cccgactatc actttgtgga ccaccgcatc gagatcctga accacgacaa
     1741 ggactacaac aaggtgaccg tttacgagag cgccgtggcc cgcaactcca ccgacggcat
     1801 ggacgagctg tacaaggggc caggtggtgc agggccaggt ggtgcagggc caggtggtgc
     1861 agggccaggt ggtgcagggc ccggtggtgc aggtccaggt ggtgcaggtc caggtggtgc
     1921 aggtccaggt ggtgctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1981 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2041 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
     2101 cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta
     2161 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2221 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2281 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2341 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
     2401 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     2461 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2521 gctgcccgac aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa
     2581 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2641 cgagctgtac aagggcggca gcggcagcga tccccccgtg gccaccatgg cgcgaccacg
     2701 gggcgcgggg ccctgcagcc ccgggttgga gcgggctccg cgccggagcg tcggggagct
     2761 gcgcctgctc ttcgaagcgc gctgcgccgc agtcgccgcc gcagcggccg cagggtaagc
     2821 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2881 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2941 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3001 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3061 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3121 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3181 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3241 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3301 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3361 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3421 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3481 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3541 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3601 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3661 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3721 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3781 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3841 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3901 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3961 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4021 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4081 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4141 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4201 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4261 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4321 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4381 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4441 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4501 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4561 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4621 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4681 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4741 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4801 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4861 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4921 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4981 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5041 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5101 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5161 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5221 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5281 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5341 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5401 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5461 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5521 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5581 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5641 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5701 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5761 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5821 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5881 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5941 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6001 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6061 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6121 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6181 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6241 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6301 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6361 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6421 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6481 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6541 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6601 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6661 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6721 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6781 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6841 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6901 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6961 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7021 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7081 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7141 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7201 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7261 gacgtc
//
