LOCUS       sequence-212951-        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"
     CDS             16..33
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             34..75
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             76..96
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(537..555)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      551..598
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(836..854)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        922..1026
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1027..1095,1096..1887)
                     /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(1245..1264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2058..2646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2547..2566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2800..2817
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2832..2971
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2897..2916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3057..3079
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3073..3264)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4576..5658)
                     /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     5620..5639
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5659..5736)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5942..5961
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6045..6064
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6045..6063
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6064..6088
                     /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             join(6103..6120,1..5)
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ggagatatac atatgcatca tcaccatcac catggtaagc ctatccctaa ccctctcctc
       61 ggtctcgatt ctacggaaaa cctgtatttt cagggaattg atcccttcac cgttgatccg
      121 attcagtttg cacgtgccaa tcaggcaatt cagatggcat gtcagaatct ggttgatccg
      181 ggtagcagcc cgagccaggt tctgagcgca gcaaccattg ttgcaaaaca taccagcgca
      241 ctgtgtaatg catgtcgtat tgcaagcagc aaaaccgcaa atccggttgc caaacgtcat
      301 tttgttcaga gcgcaaaaga agttgcaaat agcaccgcca atctggtgaa aaccattaaa
      361 gcactggatg gcgattttag cgaagataat cgtaataaat gccgcattgc aaccacaccg
      421 ctgattgaag cagttgaaaa tctgaccgca tttgcaagca atccggaatt ttaaaagggc
      481 gagctcagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
      541 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      601 gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag
      661 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
      721 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
      781 tagcttatcg atgataagct gtcaaacatg agaattaatt cttgaagacg aaagggcctc
      841 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
      901 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      961 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1021 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1081 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1141 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1201 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1261 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1321 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1381 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1441 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1501 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1561 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1621 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1681 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1741 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1801 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1861 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1921 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1981 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2701 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2761 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2821 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2881 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     2941 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     3001 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3061 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     3121 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     3181 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     3241 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     3301 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     3361 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     3421 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     3481 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     3541 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     3601 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     3661 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     3721 gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat
     3781 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa
     3841 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc
     3901 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     3961 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa
     4021 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat
     4081 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg
     4141 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc
     4201 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc
     4261 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     4321 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     4381 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     4441 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     4501 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     4561 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4621 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     4681 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     4741 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     4801 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     4861 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     4921 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     4981 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5041 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5101 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5161 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5221 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5281 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     5341 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     5401 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5461 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     5521 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     5581 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     5641 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     5701 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     5761 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     5821 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     5881 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     5941 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6001 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
     6061 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta actttaagaa
//
