LOCUS       sequence-208742-        6486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domain R13 and the dimerisation 
            domian DD..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  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 6486)
  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_R13-DD
FEATURES             Location/Qualifiers
     source          1..6486
                     /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(1187..1205)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1201..1248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1486..1504)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1572..1676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1677..1745,1746..2537)
                     /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(1895..1914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2708..3296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3197..3216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3450..3467
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3482..3621
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3547..3566)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3707..3729
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3723..3914)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5226..6308)
                     /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     6270..6289
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6309..6386)
                     /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 tcaccgatcc gaccgttatt gcagaaaccg
      421 aactgctggg tgcagcagca agcattgaag cagcagccaa aaaactggaa cagctgaaac
      481 cgcgtgcaaa accgaaacag gcagatgaaa ccctggattt tgaagaacaa attctggaag
      541 ccgcaaaaag cattgcagcc gcaaccagcg cactggttaa aagcgcaagc gcagcacagc
      601 gtgaactggt tgcacagggt aaagttggta gcattccggc aaatgcagca gatgatggtc
      661 agtggtcaca gggtctgatt agcgcagccc gtatggttgc agcagcgacc agcagcctgt
      721 gtgaagcagc caatgcaagc gttcagggtc atgcaagcga agaaaaactg attagcagcg
      781 caaaacaggt tgcagccagc accgctcagc tgctggttgc atgtaaagtt aaagcagatc
      841 aggatagcga agcaatgaaa cgtctgcagg cagcaggtaa tgcagttaaa cgtgcaagcg
      901 ataatctggt tcgtgcagca cagaaagcag catttggtaa agccgatgat gatgacgttg
      961 ttgtgaaaac caaatttgtt ggtggtatcg cacagattat tgcagcacaa gaagagatgc
     1021 tgaaaaaaga acgcgaactg gaagaagcac gtaaaaaact ggcccagatt cgtcagcagc
     1081 agtataaatt cctgccgaca gaactgcgtg aagatgaagg ttaaaagggc gagctcagat
     1141 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1201 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     1261 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     1321 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     1381 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc tagcttatcg
     1441 atgataagct gtcaaacatg agaattaatt cttgaagacg aaagggcctc gtgatacgcc
     1501 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1561 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1621 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1681 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1741 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1801 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1861 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1921 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1981 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2041 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2101 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2161 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2221 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2281 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2341 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2401 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2461 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2521 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2581 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2641 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2701 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2761 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2821 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2881 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2941 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3001 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3061 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3121 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3181 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3241 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3301 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3361 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3421 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3481 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     3541 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     3601 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3661 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3721 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     3781 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     3841 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     3901 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     3961 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     4021 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     4081 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     4141 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     4201 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     4261 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     4321 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     4381 ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
     4441 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
     4501 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg
     4561 aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg
     4621 cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga
     4681 cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct
     4741 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
     4801 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg
     4861 ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc
     4921 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     4981 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     5041 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     5101 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     5161 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     5221 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5281 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     5341 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     5401 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     5461 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     5521 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     5581 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     5641 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     5701 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     5761 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5821 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5881 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5941 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     6001 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     6061 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     6121 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     6181 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     6241 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     6301 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     6361 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     6421 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     6481 tgcatg
//
