LOCUS       sequence-214576-        6669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domains R7 and R8 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6669)
  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 6669)
  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_R7R8
FEATURES             Location/Qualifiers
     source          1..6669
                     /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(1370..1388)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1384..1431
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1669..1687)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1755..1859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1860..1928,1929..2720)
                     /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(2078..2097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2891..3479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3380..3399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3633..3650
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3665..3804
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3730..3749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3890..3912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3906..4097)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5409..6491)
                     /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     6453..6472
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6492..6569)
                     /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 aaatagcaaa gttctgggtg
      541 aaagcatggc aggtattagc cagaatgcaa aaaccggtga tctgcctgca tttggtgaat
      601 gtgttggtat tgcaagcaaa gcactgtgtg gtctgaccga agcagcagca caggcagcat
      661 atctggttgg tattagcgat ccgaatagcc aggcaggtca tcagggtctg gttgatccga
      721 ttcagtttgc acgtgccaat caggcaattc agatggcatg tcagaatctg gtggatccgg
      781 gtagcagccc gagccaggtt ctgagcgcag caaccattgt tgcaaaacat accagcgcac
      841 tgtgtaatgc atgtcgtatt gccagcagca aaaccgcaaa tccggttgca aaacgtcatt
      901 ttgttcagag cgcaaaagaa gttgcaaata gcaccgcaaa tctggtgaaa accattaaag
      961 cactggatgg cgattttagc gaagataatc gtaataaatg ccgcattgca accacaccgc
     1021 tgattgaagc agttgaaaat ctgaccgcat ttgcaagcaa tccggaattt gccagcattc
     1081 cggcacagat tagcagcgaa ggtagccagg cacaagaacc gattctggtt agcgcaaaaa
     1141 ccatgctgga aagcagcagc tatctgattc gtaccgcacg tagcctggca attaatccga
     1201 aagatccgcc tacctggtca gttctggcag gtcatagcca taccgttagc gatagcatta
     1261 aaagcctgat taccagcatt cgtgataaag ctccgggtca gcgttaaaag ggcgagctca
     1321 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     1381 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     1441 ggaactatat ccggatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
     1501 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     1561 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agctagctta
     1621 tcgatgataa gctgtcaaac atgagaatta attcttgaag acgaaagggc ctcgtgatac
     1681 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     1741 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1801 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1861 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1921 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1981 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2041 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2101 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2161 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2221 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2281 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2341 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2401 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2461 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2521 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2581 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2641 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2701 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2761 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2821 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2881 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2941 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3001 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3061 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3121 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3181 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3241 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3301 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3361 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3421 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3481 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3541 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3601 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3661 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     3721 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3781 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3841 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3901 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3961 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     4021 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     4081 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     4141 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     4201 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     4261 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4321 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4381 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4441 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4501 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     4561 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
     4621 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     4681 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     4741 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     4801 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     4861 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     4921 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     4981 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     5041 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     5101 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     5161 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     5221 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     5281 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     5341 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     5401 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5461 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     5521 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     5581 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     5641 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     5701 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     5761 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     5821 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     5881 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5941 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     6001 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     6061 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     6121 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     6181 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     6241 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     6301 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     6361 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     6421 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     6481 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     6541 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     6601 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     6661 tggtgcatg
//
