LOCUS       sequence-208733-        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     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(917..935)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      931..978
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1216..1234)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1302..1406
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1407..1475,1476..2267)
                     /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(1625..1644)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2438..3026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2927..2946
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3180..3197
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3212..3351
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3277..3296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3437..3459
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3453..3644)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4956..6038)
                     /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     6000..6019
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6039..6116)
                     /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 tcaccaaaga tgttgatgtt gccctgaaaa
      421 gcattggtga agcaagcaaa aaactgctgg ttgatagcct gcctccgagc accaaaccgt
      481 ttcaagaagc acagagcgaa ctgaatcagg cagcagcaga tctgaatcag agtgccggtg
      541 aagttgttca tgcaacccgt ggtcagagcg gtgaactggc agcagccagc ggtaaattta
      601 gtgatgattt tgatgaattc ctggacgcag gtattgaaat ggcaggtcag gcacagacca
      661 aagaagatca gatgcaggtt attggcaacc tgaaaaacat tagcatggca agtagcaaac
      721 tgctgctggc agcaaaaagc ctgagcgttg atccgggtgc accgaatgcc aaaaatctgc
      781 tggctgcagc agcccgtgca gttaccgaaa gcattaacca gctgattatg ctgtgtaccc
      841 agcaggcacc gtaaaagggc gagctcagat ccggctgcta acaaagcccg aaaggaagct
      901 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      961 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg
     1021 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     1081 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     1141 ataaactacc gcattaaagc tagcttatcg atgataagct gtcaaacatg agaattaatt
     1201 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1261 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1321 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1381 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1441 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1501 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1561 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1621 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     1681 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1741 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1801 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1861 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1921 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     1981 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2041 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2101 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2161 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2221 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2281 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2341 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2401 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2461 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2521 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2581 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2641 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2701 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2761 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2821 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2881 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2941 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3001 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3061 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3121 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3181 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3241 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3301 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3361 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3421 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3481 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     3541 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     3601 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     3661 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     3721 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     3781 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     3841 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     3901 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     3961 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4021 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4081 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc
     4141 cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc
     4201 gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta
     4261 gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca
     4321 acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg
     4381 tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc
     4441 tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg
     4501 acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg
     4561 ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg
     4621 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     4681 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     4741 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     4801 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     4861 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     4921 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4981 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5041 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     5101 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     5161 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     5221 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     5281 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     5341 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     5401 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     5461 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     5521 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     5581 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     5641 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     5701 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     5761 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     5821 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     5881 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     5941 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     6001 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     6061 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     6121 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     6181 gccgttgagc accgccgccg caaggaatgg tgcatg
//
