LOCUS       sequence-213043-        6156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod sub domain R2 a 4 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6156)
  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 6156)
  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_R2
FEATURES             Location/Qualifiers
     source          1..6156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     162..181
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        162..180
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    181..205
                     /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             220..242
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             253..270
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             271..312
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             313..333
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(810..828)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      824..871
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1109..1127)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1195..1299
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1300..1368,1369..2160)
                     /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(1518..1537)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2331..2919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2820..2839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3073..3090
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3105..3244
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3170..3189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3330..3352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3346..3537)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4849..5931)
                     /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     5893..5912
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5932..6009)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
       61 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
      121 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
      181 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
      241 gatatacata tgcatcatca ccatcaccat ggtaagccta tccctaaccc tctcctcggt
      301 ctcgattcta cggaaaacct gtattttcag ggaattgatc ccttcaccgg tgaaaacgaa
      361 accgatgaac gttttcagga tgttctgatg agcctggcaa aagcagttgc aaatgcagca
      421 gcaatgctgg ttctgaaagc aaaaaatgtt gcacaggttg cagaagatac cgtactgcag
      481 aatcgtgtta ttgcagcagc cacccagtgt gcactgagca ccagccagct ggttgcatgt
      541 gcaaaagttg ttagcccgac cattagcagt ccggtttgtc aagaacagct gattgaagca
      601 ggtaaactgg ttgatcgtag cgttgaaaat tgtgttcgtg catgtcaggc agcaaccggt
      661 gatagcgaac tgctgaaaca ggttagcgca gccgcaagcg ttgtgagcca ggcactgcat
      721 gatctgctgc aacatgttcg tcagtaaaag ggcgagctca gatccggctg ctaacaaagc
      781 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
      841 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
      901 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg
      961 gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag
     1021 caatttaact gtgataaact accgcattaa agctagctta tcgatgataa gctgtcaaac
     1081 atgagaatta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     1141 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1201 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1261 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1321 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1381 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1441 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1501 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     1561 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1621 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1681 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1741 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1801 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     1861 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1921 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1981 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2041 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2101 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2161 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2221 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2281 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2341 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2401 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2461 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2521 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2581 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2641 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2701 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2761 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2821 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2881 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2941 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3001 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3061 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3121 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     3181 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3241 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3301 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3361 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     3421 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     3481 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     3541 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     3601 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     3661 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     3721 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     3781 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     3841 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     3901 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     3961 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     4021 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     4081 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     4141 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     4201 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     4261 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
     4321 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     4381 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     4441 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
     4501 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     4561 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     4621 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     4681 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     4741 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     4801 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     4861 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4921 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     4981 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     5041 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     5101 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     5161 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     5221 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5281 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5341 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5401 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5461 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5521 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5581 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5641 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5701 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5761 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5821 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5881 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     5941 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     6001 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     6061 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     6121 cgcccaacag tcccccggcc acggggcctg ccacca
//
