LOCUS       sequence-266796-        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     151..170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     404..421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    436..575
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(501..520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     661..683
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(677..868)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2180..3262)
                     /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     3224..3243
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3263..3340)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3546..3565
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3649..3668
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3649..3667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3668..3692
                     /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             3707..3729
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3740..3757
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3758..3799
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3800..3820
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(4411..4429)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4425..4472
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4710..4728)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4796..4900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4901..4969,4970..5761)
                     /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(5119..5138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5932..6270,1..250)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
       61 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      421 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
      481 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
      541 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
      601 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
      661 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
      721 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
      781 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
      841 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
      901 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
      961 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     1021 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     1081 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     1141 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     1201 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     1261 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     1321 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     1381 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     1441 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     1501 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     1561 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     1621 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     1681 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     1741 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     1801 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     1861 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     1921 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     1981 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     2041 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     2101 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     2161 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2221 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     2281 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     2341 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     2401 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     2461 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     2521 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     2581 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     2641 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     2701 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     2761 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     2821 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     2881 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     2941 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     3001 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     3061 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     3121 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     3181 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     3241 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     3301 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     3361 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     3421 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     3481 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     3541 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     3601 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca
     3661 ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta
     3721 agaaggagat atacatatgc atcatcacca tcaccatggt aagcctatcc ctaaccctct
     3781 cctcggtctc gattctacgg aaaacctgta ttttcaggga attgatccct tcaccgcacc
     3841 gggtcagaaa gaatgtgata atgcactgcg tgaactggaa accgttaaag gtatgctgga
     3901 aaatccgaat gaaccggtta gcgatctgag ctattttgat tgtattgaga gcgtgatgga
     3961 aaacagcaaa gttctgggtg aaagcatggc aggtattagc cagaatgcaa aaaccggtga
     4021 tctgcctgca tttggtgaat gtgttggtat tgcaagcaaa gcactgtgtg gtctgaccga
     4081 agcagcagca caggcagcat atctggttgg tattagcgat ccgaatcaga ttagcagcga
     4141 aggtagccag gcacaagaac cgattctggt tagcgcaaaa accatgctgg aatcaagcag
     4201 ctatctgatc cgtaccgcac gtagcctggc aattaatccg aaagatccgc ctacctggtc
     4261 agttctggca ggtcatagcc ataccgttag cgatagcatt aaaagcctga ttaccagcat
     4321 tcgtgataaa gctccgggtc agcgttaaaa gggcgagctc agatccggct gctaacaaag
     4381 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     4441 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     4501 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     4561 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     4621 gcaatttaac tgtgataaac taccgcatta aagctagctt atcgatgata agctgtcaaa
     4681 catgagaatt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4741 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4801 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4861 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4921 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4981 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5041 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5101 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     5161 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5221 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5281 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5341 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5401 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5461 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5521 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5581 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5641 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5701 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5761 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5821 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5881 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5941 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6001 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6061 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6121 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6181 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6241 gtcgggctga acggggggtt cgtgcacaca
//
