LOCUS       sequence-208734-        6270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domain R7 a 5 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6270)
  AUTHORS   Bouchet BP, Gough RE, Ammon YC, van de Willige D, Post H, Jacquemet 
            G, Altelaar AM, Heck AJ, Goult BT, Akhmanova A
  TITLE     Talin-KANK1 interaction controls the recruitment of cortical 
            microtubule stabilizing complexes to focal adhesions.
  JOURNAL   Elife. 2016 Jul 13;5. doi: 10.7554/eLife.18124.
  PUBMED    27410476
REFERENCE   2  (bases 1 to 6270)
  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: "Elife."; 
            date: "2016-07-13"; volume: "5. doi"; pages: " 10.7554/eLife.18124"
COMMENT     Sequence Label: mTalin2_R7
FEATURES             Location/Qualifiers
     source          1..6270
                     /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(971..989)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      985..1032
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1270..1288)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1356..1460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1461..1529,1530..2321)
                     /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(1679..1698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2492..3080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2981..3000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3234..3251
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3266..3405
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3331..3350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3491..3513
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3507..3698)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5010..6092)
                     /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     6054..6073
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6093..6170)
                     /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 tcaccgcacc gggtcagaaa gaatgtgata
      421 atgcactgcg tgaactggaa accgttaaag gtatgctgga aaatccgaat gaaccggtta
      481 gcgatctgag ctattttgat tgtattgaga gcgtgatgga aaacagcaaa gttctgggtg
      541 aaagcatggc aggtattagc cagaatgcaa aaaccggtga tctgcctgca tttggtgaat
      601 gtgttggtat tgcaagcaaa gcactgtgtg gtctgaccga agcagcagca caggcagcat
      661 atctggttgg tattagcgat ccgaatcaga ttagcagcga aggtagccag gcacaagaac
      721 cgattctggt tagcgcaaaa accatgctgg aatcaagcag ctatctgatc cgtaccgcac
      781 gtagcctggc aattaatccg aaagatccgc ctacctggtc agttctggca ggtcatagcc
      841 ataccgttag cgatagcatt aaaagcctga ttaccagcat tcgtgataaa gctccgggtc
      901 agcgttaaaa gggcgagctc agatccggct gctaacaaag cccgaaagga agctgagttg
      961 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1021 aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta
     1081 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
     1141 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
     1201 taccgcatta aagctagctt atcgatgata agctgtcaaa catgagaatt aattcttgaa
     1261 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1321 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1681 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1981 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     2041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2161 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2341 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2401 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3121 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3181 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3241 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3301 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3361 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3421 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3481 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3541 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3601 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3661 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3721 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3781 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     3841 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     3901 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     3961 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4021 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4081 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4141 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     4201 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     4261 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     4321 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     4381 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4441 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     4501 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4561 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4621 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4681 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4741 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4801 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4861 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4921 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4981 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5101 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5161 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5221 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5281 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5341 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5401 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5461 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5521 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5581 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5641 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5701 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     5761 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     5821 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     5881 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     5941 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6001 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6061 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6121 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6181 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6241 gagcaccgcc gccgcaagga atggtgcatg
//
