LOCUS       Exported                6237 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Plasmid expresses the talin2 rod domain R10 a 5 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    mTalin2_R10
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6237)
  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 6237)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6237
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     69..88
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     172..191
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        172..190
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    191..215
                     /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             230..252
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             263..280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             281..322
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             323..343
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(901..919)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      915..962
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1200..1218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1286..1390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1391..2251
                     /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(1609..1628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2422..3010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2911..2930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3164..3181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3196..3335
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3261..3280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3421..3443
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3437..3628)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4940..6022)
                     /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     5984..6003
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6023..6100)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
       61 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
      121 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
      181 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
      241 ttaagaagga gatatacata tgcatcatca ccatcaccat ggtaagccta tccctaaccc
      301 tctcctcggt ctcgattcta cggaaaacct gtattttcag ggaattgatc ccttcaccaa
      361 tgaagcagca agcgaagttg gtctggttgg tggtatggtt gatgcaattg ccgaagcaat
      421 gagcaaactg gatgaaggca cccctccgga accgaaaggc acctttgttg attatcagac
      481 caccgttgtg aaatacagca aagcaattgc agttaccgca caagaaatga tgaccaaaag
      541 cgttaccaat ccggaagaac tgggtggtct ggcaagccag atgaccaccg attatggcca
      601 tctggcactg cagggtcaaa tggcagcagc aaccgcagaa ccggaagaaa ttggttttca
      661 gattcgtacc cgtgttcagg atctgggtca tggttgtatt tttctggttc agaaagccgg
      721 tgccctgcag gtttgtccga ccgatagcta taccaaacgt gaactgattg aatgtgcacg
      781 tagcgttacc gaaaaagtta gcctggttct gagtgcgctg caggcaggta acaaataaaa
      841 gggcgagctc agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
      901 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      961 tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac
     1021 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     1081 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     1141 aagctagctt atcgatgata agctgtcaaa catgagaatt aattcttgaa gacgaaaggg
     1201 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1381 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1501 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1561 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1621 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1681 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1801 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1921 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1981 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2221 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2281 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2341 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3061 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3181 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3241 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3301 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3361 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3421 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3481 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3541 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3601 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3661 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     3721 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     3781 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     3841 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     3901 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     3961 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4021 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4081 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     4141 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     4201 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     4261 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     4321 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     4381 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     4441 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     4501 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     4561 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     4621 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4681 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4741 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4801 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     4861 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     4921 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4981 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     5041 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     5101 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     5161 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     5221 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     5281 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     5341 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     5401 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     5461 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     5521 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     5581 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     5641 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     5701 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     5761 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     5821 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     5881 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     5941 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     6001 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     6061 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     6121 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     6181 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacgggg
//
