LOCUS       sequence-375050-        4991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses human talin R7R8 domain flanked between a HaloTag 
            and a AviTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  AUTHORS   Dahal N, Sharma S, Phan B, Eis A, Popa I
  TITLE     Mechanical regulation of talin through binding and history-dependent
            unfolding.
  JOURNAL   Sci Adv. 2022 Jul 15;8(28):eabl7719. doi: 10.1126/sciadv.abl7719. 
            Epub 2022 Jul 15.
  PUBMED    35857491
REFERENCE   2  (bases 1 to 4991)
  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: "Sci Adv."; 
            date: "2022-07-15"; volume: "8(28):eabl7719. doi"; pages: " 
            10.1126/sciadv.abl7719. Epub 2022 Jul 15"
COMMENT     Sequence Label: pFN-HaloTag-TalinR7R8-AviTag
FEATURES             Location/Qualifiers
     source          1..4991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      47..74
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     156..175
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        156..174
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             205..1095
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1108..1128
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2068..2085
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2092..2136
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2230..2248)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2244..2291
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2520..2624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2625..2693,2694..3485)
                     /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(2843..2862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3643..4231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3723..3742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4347..4630
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     primer_bind     complement(4690..4707)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4860..4946
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
       61 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
      121 tatgtatccg ctcatggatc tcgatcccgc gaaattaata cgactcacta tagggtattt
      181 taactttact aaggagaatt catcatggca gaaatcggta ctggctttcc attcgacccc
      241 cattatgtgg aagtcctggg cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc
      301 acccctgtgc tgttcctgca cggtaacccg acctcctcct acgtgtggcg caacatcatc
      361 ccgcatgttg caccgaccca tcgctgcatt gctccagacc tgatcggtat gggcaaatcc
      421 gacaaaccag acctgggtta tttcttcgac gaccacgtcc gcttcatgga tgccttcatc
      481 gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg actggggctc cgctctgggt
      541 ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat ggagttcatc
      601 cgccctatcc cgacctggga cgaatggcca gaatttgccc gcgagacctt ccaggccttc
      661 cgcaccaccg acgtcggccg caagctgatc atcgatcaga acgtttttat cgagggtacg
      721 ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga tggaccatta ccgcgagccg
      781 ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc caaacgagct gccaatcgcc
      841 ggtgagccag cgaacatcgt cgcgctggtc gaagaataca tggactggct gcaccagtcc
      901 cctgtcccga agctgctgtt ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc
      961 gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg acatcggccc gggtctgaat
     1021 ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg gctgtcgacg
     1081 ctcgagattt ccggcgagcc aaccactgag gatctgtact ttcagagcga taacgcgatc
     1141 gccggatcca tgcagcaggc accgggccaa aaagaatgtg acaatgccct gcgcgaactg
     1201 gagaccgtgc gcgaactgct ggagaatccg gtgcagccga tcaatgatat gagttatttc
     1261 ggctgcctgg acagtgtgat ggagaacagc aaagtgctgg gtgaggccat gaccggtatc
     1321 agccagaacg ccaaaaacgg caatctgccg gaattcggcg atgccatcag cacagcaagc
     1381 aaagccctgt gcggctttac cgaagcagcc gcccaggcag catatctggt gggtgtgagc
     1441 gatccgaata gccaggccgg tcaacagggt ctggttgaac cgacccagtt tgcccgcgcc
     1501 aatcaggcca tccagatggc atgccagagc ttaggtgagc cgggttgcac ccaggcccag
     1561 gttctgagcg ccgcaaccat tgttgccaaa cataccagcg ccctgtgtaa tagctgccgc
     1621 ctggcaagcg cccgtaccac aaatccgacc gccaaacgcc agttcgtgca gagcgccaaa
     1681 gaagtggcca atagcacagc caacctggtt aaaaccatca aggccctgga cggtgcattt
     1741 accgaggaaa atcgcgcaca gtgtcgcgca gcaacagccc cgctgctgga agcagttgac
     1801 aacctgagcg ccttcgccag caatcctgag tttagcagca ttccggcaca gattagcccg
     1861 gagggccgcg cagcaatgga gccgattgtg atcagcgcca aaacaatgct ggaaagcgcc
     1921 ggcggcttaa tccagacagc ccgtgccctg gccgtgaatc ctcgcgatcc gccgagctgg
     1981 agtgttctgg ccggtcatag ccgtaccgtg agcgacagca tcaagaagct gatcacaagc
     2041 atgcgcgata aagcaccggg tccatggcat catcatcatc accacggcgg cggcttaaac
     2101 gacatctttg aagcacagaa aattgaatgg catgaataag gtaccagatc ctctagagtc
     2161 gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     2221 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     2281 ggggtttttt gctgaaagga ggaactatat ccggttcgct tgctgtccat aaaaccgccc
     2341 agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt
     2401 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
     2461 gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2521 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2581 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2641 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2701 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2761 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2821 tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2881 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2941 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3001 ccatgagtga taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc
     3061 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3121 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3181 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3241 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3301 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3361 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3421 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3481 ggtaattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     3541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3601 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3661 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3721 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3781 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3841 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3901 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3961 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4021 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4081 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4141 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4201 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     4261 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4321 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
     4381 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
     4441 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
     4501 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
     4561 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
     4621 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
     4681 tcttttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     4741 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     4801 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     4861 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     4921 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     4981 ggcccggagg g
//
