LOCUS Exported 6243 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Plasmid expresses the talin2 rod domain R5 a 5 helix bundle.. ACCESSION . VERSION . KEYWORDS mTalin2_R5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6243) 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 6243) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6243 /organism="synthetic DNA construct" /mol_type="other DNA" 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(944..962) /label=T7 Term /note="T7 terminator, reverse primer" terminator 958..1005 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(1243..1261) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1329..1433 /gene="bla" /label=AmpR promoter CDS 1434..2294 /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(1652..1671) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2465..3053 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2954..2973 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3207..3224 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3239..3378 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3304..3323) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3464..3486 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3480..3671) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4983..6065) /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 6027..6046 /label=LacI-R /note="LacI, reverse primer" promoter complement(6066..6143) /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 tcaccccgat ggaaattgat agcgcactga 421 ataccgttca gaccctgaaa aatgaactgc aggatgcaaa aatggcagca gcagaaagcc 481 agctgaaacc gctgcctggt gaaacactgg aaaaatgtgc acaggatctg ggtagcacca 541 gcaaaggtgt tggtagcagc atggcacagc tgctgacctg tgcagcccag ggtaatgaac 601 actataccgg tgttgcagca cgtgaaaccg cacaggcact gaaaaccctg gcacaggcag 661 cccgtggtgt tgccgcaagc accaatgatc cggaagcagc acatgcaatg ctggatagtg 721 cacgtgatgt tatggaaggt agcgcaatgc tgattcaaga agcaaaacag gccctgattg 781 caccgggtga taccgaaagt cagcagcgtc tggcccaggt tgcaaaagca gttagccata 841 gcctgaataa ttgtgttaat tgtctgcctg gccagaaata aaagggcgag ctcagatccg 901 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta 961 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact 1021 atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag 1081 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg 1141 cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctag cttatcgatg 1201 ataagctgtc aaacatgaga attaattctt gaagacgaaa gggcctcgtg atacgcctat 1261 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg 1321 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 1381 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 1441 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 1501 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 1561 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 1621 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg 1681 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 1741 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 1801 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 1861 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 1921 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag 1981 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 2041 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 2101 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 2161 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 2221 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 2281 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 2341 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 2401 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 2461 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 2521 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 2581 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 2641 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 2701 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 2761 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 2821 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 2881 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 2941 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 3001 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 3061 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 3121 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 3181 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 3241 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct 3301 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt 3361 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct 3421 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt 3481 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg 3541 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc 3601 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc 3661 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg 3721 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg 3781 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact 3841 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag 3901 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga 3961 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg 4021 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg 4081 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc 4141 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg 4201 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct 4261 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg 4321 tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc 4381 ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga 4441 tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc 4501 cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc 4561 cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca 4621 gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga 4681 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata 4741 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga 4801 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg 4861 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca 4921 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc 4981 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 5041 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt 5101 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg 5161 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata 5221 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc 5281 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc 5341 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg 5401 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta 5461 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg 5521 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg 5581 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca 5641 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc 5701 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg 5761 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc 5821 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc 5881 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc 5941 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg 6001 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt 6061 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag 6121 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat 6181 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc 6241 atg //