LOCUS       sequence-212950-        6261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domain R11 a 5 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6261)
  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 6261)
  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_R11
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..27
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             28..69
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             70..90
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(672..690)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      686..733
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(971..989)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1057..1161
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1162..1230,1231..2022)
                     /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(1380..1399)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2193..2781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2682..2701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2935..2952
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2967..3106
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3032..3051)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3192..3214
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3208..3399)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4711..5793)
                     /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     5755..5774
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5794..5871)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6077..6096
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6180..6199
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6180..6198
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6199..6223
                     /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             6238..6260
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atacatatgc atcatcacca tcaccatggt aagcctatcc ctaaccctct cctcggtctc
       61 gattctacgg aaaacctgta ttttcaggga attgatccct tcaccggcac ccaggcatgt
      121 attaccgcag caaccgcagt tagcggtatt attgcagatc tggataccac cattatgttt
      181 gccaccgcag gcaccctgaa tgcagaaaat ggtgaaacct ttgcagatca ccgtgaaaac
      241 attctgaaaa ccgcaaaagc actggtggaa gataccaaac tgctggttag cggtgcagca
      301 agcacaccgg ataaactggc acaggcagca cagagcagcg cagccaccat tacccagctg
      361 gccgaagttg ttaaactggg tgcagccagc ctgggtagca atgatccgga aacccaggtt
      421 gttctgatta acgcaattaa agatgttgca aaagccctga gcgatctgat tggtgcaacc
      481 aaaggtgcag catcaaaacc ggcagatgat ccgagcatgt atcagctgaa aggtgccgca
      541 aaagttatgg ttaccaatgt taccagcctg ctgaaaacag ttaaagccgt tgaagatgaa
      601 gccacctaaa agggcgagct cagatccggc tgctaacaaa gcccgaaagg aagctgagtt
      661 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
      721 gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag gcccggcagt
      781 accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg
      841 agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa
      901 ctaccgcatt aaagctagct tatcgatgat aagctgtcaa acatgagaat taattcttga
      961 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1021 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1081 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1141 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1201 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1261 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1321 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1381 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     1441 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1501 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1561 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1621 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1681 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     1741 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1801 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1861 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1921 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1981 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2041 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2101 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2161 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2221 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2281 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2341 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2401 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2461 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2521 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2581 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2641 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2701 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2761 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2821 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2881 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2941 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3001 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3061 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3121 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3181 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3241 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     3301 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     3361 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     3421 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     3481 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     3541 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     3601 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     3661 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     3721 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     3781 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     3841 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt
     3901 gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt
     3961 cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag
     4021 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
     4081 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
     4141 gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta
     4201 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
     4261 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
     4321 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
     4381 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     4441 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     4501 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     4561 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     4621 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     4681 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4741 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4801 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     4861 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     4921 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     4981 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5041 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5101 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5161 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5221 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     5281 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     5341 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     5401 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     5461 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     5521 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     5581 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     5641 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     5701 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     5761 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     5821 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     5881 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     5941 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6001 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6061 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6121 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     6181 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     6241 gtttaacttt aagaaggaga t
//
