LOCUS       sequence-214611-        6135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod sub domain R3 a 4 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6135)
  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 6135)
  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_R3
FEATURES             Location/Qualifiers
     source          1..6135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    115..254
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(180..199)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     340..362
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(356..547)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1859..2941)
                     /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     2903..2922
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2942..3019)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3225..3244
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3328..3347
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3328..3346
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3347..3371
                     /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             3386..3408
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3419..3436
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3437..3478
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3479..3499
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3955..3973)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3969..4016
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4254..4272)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4340..4444
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4445..4513,4514..5305)
                     /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(4663..4682)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5476..6064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5965..5984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
       61 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      121 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      181 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      241 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      301 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
      361 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
      421 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
      481 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
      541 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
      601 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
      661 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
      721 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
      781 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
      841 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
      901 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
      961 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     1021 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
     1081 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     1141 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     1201 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     1261 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     1321 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     1381 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     1441 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     1501 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     1561 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     1621 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     1681 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     1741 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     1801 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     1861 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1921 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     1981 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     2041 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     2101 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     2161 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     2221 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     2281 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     2341 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     2401 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     2461 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     2521 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     2581 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     2641 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     2701 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     2761 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     2821 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     2881 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     2941 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     3001 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     3061 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     3121 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     3181 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     3241 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     3301 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     3361 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgca
     3421 tcatcaccat caccatggta agcctatccc taaccctctc ctcggtctcg attctacgga
     3481 aaacctgtat tttcagggaa ttgatccctt cacctttgca agccgtggtg aaccgattgg
     3541 tcgttatgat caggcaaccg ataccattat gtgtgttacc gaaagcattt ttagcagcat
     3601 gggtgatgcc ggtgaaatgg ttcgtcaggc acgtgttctg gcacaggcaa ccagcgatct
     3661 ggttaatgca atgcgtagtg atgccgaagc agaaattgat atggaaaaca gcaaaaaact
     3721 gctggcagca gccaaactgc tggccgatag caccgcacgt atggttgaag cagcaaaagg
     3781 tgcagcagca aatccggaaa atgaagatca gcagcagcgt ctgcgtgaag cagccgaagg
     3841 tctgcgtgtt gcaaccaatg cagcagccca gaatgcaatc aaaaaaaaat aaaagggcga
     3901 gctcagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     3961 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     4021 aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
     4081 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
     4141 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagcta
     4201 gcttatcgat gataagctgt caaacatgag aattaattct tgaagacgaa agggcctcgt
     4261 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4321 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4381 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4441 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4501 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4561 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4621 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4681 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4741 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4801 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4861 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4921 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4981 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5041 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5101 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5161 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5221 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5281 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5341 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5401 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5461 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5521 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5581 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5641 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5701 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5761 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5821 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5881 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5941 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6001 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6061 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6121 catgttcttt cctgc
//
