LOCUS Exported 6111 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Plasmid expresses mouse talin2 FERM domain F1. ACCESSION . VERSION . KEYWORDS mTalin2_F1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6111) 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 6111) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6111 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 96..115 /label=T7 /note="T7 promoter, forward primer" promoter 96..114 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 115..139 /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 154..176 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 187..204 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 205..246 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 247..267 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" primer_bind complement(699..717) /label=T7 Term /note="T7 terminator, reverse primer" terminator 713..760 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(998..1016) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1084..1188 /gene="bla" /label=AmpR promoter CDS 1189..2049 /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(1407..1426) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2220..2808 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2709..2728 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2962..2979 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2994..3133 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3059..3078) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3219..3241 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3235..3426) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4738..5820) /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 5782..5801 /label=LacI-R /note="LacI, reverse primer" promoter complement(5821..5898) /gene="lacI" /label=lacI promoter primer_bind 6104..12 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc 61 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt 121 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata 181 catatgcatc atcaccatca ccatggtaag cctatcccta accctctcct cggtctcgat 241 tctacggaaa acctgtattt tcagggaatt gatcccttca cccgtccgca gaaaattcgt 301 atgctggatg gtagcgttaa aaccgttatg gttgatgata gcaaaaccgt tggtgaactg 361 ctggttacca tttgtagccg tattggcatt accaactatg aagaatatag cctgatccaa 421 gaaaccatcg aagaaaaaaa agaagaaggc accggcaccc tgaaaaaaga tcgtaccctg 481 ctgcgtgatg aacgcaaaat ggaaaaactg aaagcaaaac tgcacaccga tgatgatctg 541 aattggctgg atcatagccg tacctttcgt gaacagggtg ttgatgaaaa tgaaacactg 601 ctgctgcgtc gcaaattttt ctatagcgat cagtaaaagg gcgagctcag atccggctgc 661 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata 721 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc 781 cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc 841 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga 901 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gctagcttat cgatgataag 961 ctgtcaaaca tgagaattaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta 1021 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat 1081 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 1141 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 1201 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 1261 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 1321 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 1381 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc 1441 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 1501 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 1561 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 1621 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 1681 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg 1741 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 1801 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 1861 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 1921 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 1981 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 2041 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 2101 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 2161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 2221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 2281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 2341 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc 2401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 2461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 2521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 2581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 2641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 2701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 2761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 2821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 2881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 2941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg 3001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta 3061 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 3121 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3241 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 3301 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 3361 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 3421 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 3481 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 3541 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 3601 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 3661 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3721 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3781 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3841 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga 3901 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat 3961 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat 4021 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc 4081 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca 4141 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc 4201 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga 4261 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg 4321 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag 4381 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt 4441 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca 4501 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag 4561 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg 4621 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc 4681 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca 4741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 4801 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac 4861 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac 4921 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca 4981 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc 5041 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc 5101 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact 5161 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca 5221 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg 5281 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa 5341 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt 5401 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc 5461 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg 5521 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc 5581 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga 5641 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc 5701 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga 5761 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac 5821 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt 5881 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga 5941 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca 6001 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac 6061 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg t //