LOCUS       sequence-208725-        6105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses mouse talin2 FERM domain F2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6105)
  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 6105)
  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_F2
FEATURES             Location/Qualifiers
     source          1..6105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             300..317
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..359
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             360..380
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(806..824)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      820..867
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1105..1123)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1191..1295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1296..1364,1365..2156)
                     /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(1514..1533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2327..2915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2816..2835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3069..3086
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3101..3240
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3166..3185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3326..3348
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3342..3533)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4845..5927)
                     /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     5889..5908
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5928..6005)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgc
      301 atcatcacca tcaccatggt aagcctatcc ctaaccctct cctcggtctc gattctacgg
      361 aaaacctgta ttttcaggga attgatccct tcaccaaatt cttctacagc gatcagaatg
      421 tggatagccg tgatccggtg cagctgaatc tgctgtatgt tcaggcacgt gatgatattc
      481 tgaatggtag ccatccggtt agctttgaaa aagcatgtga atttggtggt tttcaggccc
      541 agattcagtt tggtccgcat gttgaacata aacacaaacc gggttttctg gatctgaaag
      601 aatttctgcc gaaagaatac attaaacagc gtggtgcaga aaaacgcatc tttcaagaac
      661 ataaaaactg cggtgaaatg agcgaaattg aggccaaagt gaaatatgtt aaactggcac
      721 gtagcctgcg tacctatggt taaaagggcg agctcagatc cggctgctaa caaagcccga
      781 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      841 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatcccgca
      901 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
      961 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
     1021 ttaactgtga taaactaccg cattaaagct agcttatcga tgataagctg tcaaacatga
     1081 gaattaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     1141 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1201 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1261 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1321 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1381 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1441 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1501 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     1561 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1621 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1681 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1741 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1801 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     1861 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1921 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1981 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2041 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2101 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2161 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2221 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2281 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2341 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2401 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2461 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2521 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2581 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2641 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2701 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2761 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2821 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2881 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2941 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3001 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3061 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3121 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     3181 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     3241 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3301 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3361 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     3421 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     3481 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     3541 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     3601 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     3661 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     3721 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     3781 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     3841 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     3901 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     3961 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     4021 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg
     4081 ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg
     4141 aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac
     4201 cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc
     4261 cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg
     4321 aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta
     4381 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     4441 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     4501 cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     4561 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     4621 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     4681 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     4741 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     4801 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     4861 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4921 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     4981 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     5041 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     5101 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     5161 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     5221 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     5281 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     5341 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     5401 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     5461 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     5521 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     5581 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     5641 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     5701 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     5761 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     5821 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     5881 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     5941 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     6001 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     6061 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatg
//
