LOCUS       sequence-212972-        6039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses mouse talin2 FERM domain F3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  AUTHORS   Gough RE, Goult BT
  TITLE     The tale of two talins - two isoforms to fine-tune integrin 
            signalling.
  JOURNAL   FEBS Lett. 2018 May 3. doi: 10.1002/1873-3468.13081.
  PUBMED    29723415
REFERENCE   2  (bases 1 to 6039)
  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: "FEBS Lett. 
            2018 May 3. doi: 10.1002/1873-3468.13081."
COMMENT     Sequence Label: mTalin2_F3
FEATURES             Location/Qualifiers
     source          1..6039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     124..143
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        124..142
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    143..167
                     /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)."
     RBS             182..204
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             215..232
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             233..274
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             275..295
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(655..673)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      669..716
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(954..972)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1040..1144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1145..1213,1214..2005)
                     /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     complement(1363..1382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2176..2764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2665..2684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2918..2935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2950..3089
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3015..3034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3175..3197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3191..3382)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4694..5776)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5738..5757
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5777..5854)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
       61 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
      121 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
      181 ttttgtttaa ctttaagaag gagatataca tatgcatcat caccatcacc atggtaagcc
      241 tatccctaac cctctcctcg gtctcgattc tacggaaaac ctgtattttc agggaattga
      301 tcccttcacc tatggcgtga gcttttttct ggtgaaagaa aaaatgaaag gcaaaaacaa
      361 actggttccg cgtctgctgg gtattaccaa agatagcgtt atgcgtgttg acgagaaaac
      421 caaagaagtt ctgcaagaat ggcctctgac caccgttaaa cgttgggcag caagcccgaa
      481 aagctttacc ctggattttg gtgaatatca agagtcctat tattcagtgc agaccaccga
      541 aggtgagcag attagccagc tgattgcagg ttatattgat atcatcctgt aaaagggcga
      601 gctcagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      661 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      721 aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
      781 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
      841 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagcta
      901 gcttatcgat gataagctgt caaacatgag aattaattct tgaagacgaa agggcctcgt
      961 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     1021 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1081 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1141 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1201 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1261 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1321 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1381 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1441 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1501 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1561 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1621 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1681 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1741 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1801 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1861 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1921 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1981 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2041 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2101 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2161 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2221 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2281 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2341 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2401 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2461 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2521 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2581 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2641 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2701 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2761 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2821 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2881 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2941 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     3001 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3061 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3121 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3181 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3241 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3301 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3361 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3421 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3481 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     3541 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     3601 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     3661 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     3721 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     3781 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     3841 gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg
     3901 cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga
     3961 attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat
     4021 tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta
     4081 tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat
     4141 cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc
     4201 ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg
     4261 catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt
     4321 cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg
     4381 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     4441 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     4501 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     4561 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     4621 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     4681 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4741 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4801 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4861 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4921 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     4981 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     5041 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     5101 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     5161 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     5221 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     5281 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     5341 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     5401 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     5461 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     5521 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     5581 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     5641 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     5701 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5761 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     5821 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5881 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5941 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     6001 atacccacgc cgaaacaagc gctcatgagc ccgaagtgg
//
