LOCUS       sequence-212959-        6282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domain R1 a 5 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6282)
  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 6282)
  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_R1
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     151..170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        151..169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    170..194
                     /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             209..231
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             242..259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..301
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             302..322
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(925..943)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      939..986
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1224..1242)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1310..1414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1415..1483,1484..2275)
                     /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(1633..1652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2446..3034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2935..2954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3188..3205
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3220..3359
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3285..3304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3445..3467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3461..3652)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4964..6046)
                     /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     6008..6027
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6047..6124)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
       61 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
      121 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
      181 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat
      241 gcatcatcac catcaccatg gtaagcctat ccctaaccct ctcctcggtc tcgattctac
      301 ggaaaacctg tattttcagg gaattgatcc cttcaccggt catatgcctc cgctgaccag
      361 cgcacagcag gcactgatgg gcaccattaa taccagtatg catgcagttc agcaggccca
      421 ggatgatctg agcgaactgg atagcctgcc accgctgggt caggatatgg caagccgtgt
      481 ttgggttcag aataaagttg atgaaagcaa acacgaaatc cacagccagg ttgatgcaat
      541 taccgcaggc accgcaagcg ttgttaatct gaccgcaggc gatccggcag ataccgatta
      601 taccgcagtt ggttgtgcca ttaccaccat tagcagcaat ctgacagaaa tgagcaaagg
      661 tgttaaactg ctggcagccc tgatggatga tgatgttggt agcggtgaag atctgctgcg
      721 tgcagcacgt accctggcag gcgcagttag cgatctgctg aaagccgttc agccgaccag
      781 cggtgaaccg cgtcagaccg ttctgacagc agcaggtagc attggtcagg caagcggtga
      841 cctgctgcgc cagattggtt aaaagggcga gctcagatcc ggctgctaac aaagcccgaa
      901 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      961 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa
     1021 gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc
     1081 gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt
     1141 taactgtgat aaactaccgc attaaagcta gcttatcgat gataagctgt caaacatgag
     1201 aattaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1261 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1321 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1381 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1441 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1501 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1561 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1621 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     1681 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1741 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1801 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1861 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1921 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     1981 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2041 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2101 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2161 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2221 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2281 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2341 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2401 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2461 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2521 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2581 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2641 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2701 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2761 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2821 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2881 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2941 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3001 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3061 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3121 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3181 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     3241 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     3301 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     3361 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3421 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3481 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     3541 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     3601 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     3661 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     3721 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     3781 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     3841 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     3901 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     3961 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     4021 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     4081 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg
     4141 agatgcgccg cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt
     4201 tggtttgcgc attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga
     4261 atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc
     4321 gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc
     4381 gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga
     4441 agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac
     4501 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
     4561 cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc
     4621 gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     4681 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     4741 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     4801 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     4861 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     4921 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     4981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5041 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     5101 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     5161 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     5221 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     5281 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     5341 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     5401 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     5461 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     5521 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     5581 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     5641 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     5701 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     5761 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     5821 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     5881 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     5941 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     6001 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     6061 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     6121 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     6181 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     6241 aacagtcccc cggccacggg gcctgccacc atacccacgc cg
//
