LOCUS       sequence-212961-        6216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domain R6 a 5 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6216)
  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 6216)
  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_R6
FEATURES             Location/Qualifiers
     source          1..6216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..72
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        53..71
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    72..96
                     /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             111..133
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             144..161
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             162..203
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             204..224
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(761..779)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      775..822
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1060..1078)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1146..1250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1251..1319,1320..2111)
                     /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(1469..1488)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2282..2870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2771..2790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3024..3041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3056..3195
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3121..3140)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3281..3303
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3297..3488)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4800..5882)
                     /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     5844..5863
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5883..5960)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6166..6185
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
       61 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      121 tttaagaagg agatatacat atgcatcatc accatcacca tggtaagcct atccctaacc
      181 ctctcctcgg tctcgattct acggaaaacc tgtattttca gggaattgat cccttcacca
      241 aagatgttga tgttgccctg aaaagcattg gtgaagcaag caaaaaactg ctggttgata
      301 gcctgcctcc gagcaccaaa ccgtttcaag aagcacagag cgaactgaat caggcagcag
      361 cagatctgaa tcagagtgcc ggtgaagttg ttcatgcaac ccgtggtcag agcggtgaac
      421 tggcagcagc cagcggtaaa tttagtgatg attttgatga attcctggac gcaggtattg
      481 aaatggcagg tcaggcacag accaaagaag atcagatgca ggttattggc aacctgaaaa
      541 acattagcat ggcaagtagc aaactgctgc tggcagcaaa aagcctgagc gttgatccgg
      601 gtgcaccgaa tgccaaaaat ctgctggctg cagcagcccg tgcagttacc gaaagcatta
      661 accagctgat tatgctgtgt acccagcagg caccgtaaaa gggcgagctc agatccggct
      721 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      781 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      841 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
      901 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
      961 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagctagctt atcgatgata
     1021 agctgtcaaa catgagaatt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     1081 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1141 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1201 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1261 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1321 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1381 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1441 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     1501 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1561 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1621 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1681 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1741 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     1801 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1861 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1921 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1981 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2041 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2101 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2161 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2221 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2281 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2341 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2401 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2461 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2521 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2581 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2641 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2701 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2761 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2821 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2881 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2941 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3001 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3061 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3121 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3181 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3241 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3301 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3361 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3421 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     3481 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     3541 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     3601 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     3661 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     3721 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     3781 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     3841 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     3901 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     3961 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     4021 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     4081 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     4141 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     4201 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     4261 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     4321 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     4381 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     4441 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     4501 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     4561 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     4621 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     4681 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4741 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4801 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4861 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     4921 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     4981 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     5041 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     5101 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5161 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5221 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5281 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5341 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5401 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5461 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5521 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5581 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5641 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5701 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5761 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5821 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     5881 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     5941 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6001 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     6061 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     6121 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     6181 ataggcgcca gcaaccgcac ctgtggcgcc ggtgat
//
