LOCUS       sequence-208735-        6120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domain R8 a 4 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6120)
  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 6120)
  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_R8
FEATURES             Location/Qualifiers
     source          1..6120
                     /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(821..839)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      835..882
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1120..1138)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1206..1310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1311..1379,1380..2171)
                     /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(1529..1548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2342..2930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2831..2850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3084..3101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3116..3255
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3181..3200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3341..3363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3357..3548)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4860..5942)
                     /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     5904..5923
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5943..6020)
                     /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 tcaccgttga tccgattcag tttgcacgtg
      421 ccaatcaggc aattcagatg gcatgtcaga atctggttga tccgggtagc agcccgagcc
      481 aggttctgag cgcagcaacc attgttgcaa aacataccag cgcactgtgt aatgcatgtc
      541 gtattgcaag cagcaaaacc gcaaatccgg ttgccaaacg tcattttgtt cagagcgcaa
      601 aagaagttgc aaatagcacc gccaatctgg tgaaaaccat taaagcactg gatggcgatt
      661 ttagcgaaga taatcgtaat aaatgccgca ttgcaaccac accgctgatt gaagcagttg
      721 aaaatctgac cgcatttgca agcaatccgg aattttaaaa gggcgagctc agatccggct
      781 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      841 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      901 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
      961 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     1021 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagctagctt atcgatgata
     1081 agctgtcaaa catgagaatt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     1141 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1201 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1261 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1321 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1381 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1441 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1501 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     1561 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1621 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1681 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1741 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1801 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     1861 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1921 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1981 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2041 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2101 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2161 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2221 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2281 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2341 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2401 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2461 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2521 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2581 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2641 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2701 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2761 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2821 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2881 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2941 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3001 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3061 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3121 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3181 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3241 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3301 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3361 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3421 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3481 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     3541 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     3601 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     3661 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     3721 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     3781 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     3841 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     3901 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     3961 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     4021 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     4081 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     4141 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     4201 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     4261 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     4321 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     4381 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     4441 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     4501 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     4561 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     4621 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     4681 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     4741 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4801 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4921 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     4981 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5041 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     5101 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     5161 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5221 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5281 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5341 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5401 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5461 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5521 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5581 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5641 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5701 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5761 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5821 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5881 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     5941 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6001 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6061 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
//
