LOCUS       sequence-212956-        6105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses mouse talin2 FERM domain F2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6105)
  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 6105)
  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_F2
FEATURES             Location/Qualifiers
     source          1..6105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..102
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        83..101
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    102..126
                     /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             141..163
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             174..191
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             192..233
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             234..254
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(680..698)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      694..741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(979..997)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1065..1169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1170..1238,1239..2030)
                     /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(1388..1407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2201..2789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2690..2709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2943..2960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2975..3114
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3040..3059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3200..3222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3216..3407)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4719..5801)
                     /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     5763..5782
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5802..5879)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6085..6104
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
       61 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
      121 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat atgcatcatc
      181 accatcacca tggtaagcct atccctaacc ctctcctcgg tctcgattct acggaaaacc
      241 tgtattttca gggaattgat cccttcacca aattcttcta cagcgatcag aatgtggata
      301 gccgtgatcc ggtgcagctg aatctgctgt atgttcaggc acgtgatgat attctgaatg
      361 gtagccatcc ggttagcttt gaaaaagcat gtgaatttgg tggttttcag gcccagattc
      421 agtttggtcc gcatgttgaa cataaacaca aaccgggttt tctggatctg aaagaatttc
      481 tgccgaaaga atacattaaa cagcgtggtg cagaaaaacg catctttcaa gaacataaaa
      541 actgcggtga aatgagcgaa attgaggcca aagtgaaata tgttaaactg gcacgtagcc
      601 tgcgtaccta tggttaaaag ggcgagctca gatccggctg ctaacaaagc ccgaaaggaa
      661 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      721 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc
      781 ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga
      841 tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact
      901 gtgataaact accgcattaa agctagctta tcgatgataa gctgtcaaac atgagaatta
      961 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1021 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1141 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1201 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1261 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1321 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1381 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     1441 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1561 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1621 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1681 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     1741 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1801 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1861 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1921 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1981 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2041 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2101 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2161 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2221 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2281 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2341 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2401 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2461 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2521 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2581 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2641 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2701 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2761 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2821 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2881 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2941 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3001 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     3061 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3121 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3181 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3241 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3301 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3361 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     3421 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     3481 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     3541 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     3601 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     3661 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     3721 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     3781 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     3841 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg
     3901 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     3961 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     4021 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac
     4081 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc
     4141 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta
     4201 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc
     4261 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     4321 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     4381 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     4441 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4501 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4561 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4621 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4681 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4741 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4801 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     4861 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4921 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     4981 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5041 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5101 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5161 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5221 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     5281 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     5341 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5401 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     5461 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     5521 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     5581 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     5641 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     5701 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     5761 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     5821 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     5881 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     5941 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     6001 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     6061 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggc
//
