LOCUS       sequence-362821-        6390 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses mouse Talin 1 F2F3-P229S domains in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6390)
  AUTHORS   Azizi L, Varela L, Turkki P, Mykuliak VV, Korpela S, Ihalainen TO, 
            Church J, Hytonen VP, Goult BT
  TITLE     Talin variant P229S compromises integrin activation and associates 
            with multifaceted clinical symptoms.
  JOURNAL   Hum Mol Genet. 2022 Jul 21. pii: 6647920. doi: 10.1093/hmg/ddac163.
  PUBMED    35861643
REFERENCE   2  (bases 1 to 6390)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Hum Mol 
            Genet. 2022 Jul 21. pii: 6647920. doi: 10.1093/hmg/ddac163."
COMMENT     Sequence Label: mTln1-F2F3-P229S-pET151
FEATURES             Location/Qualifiers
     source          1..6390
                     /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(1091..1109)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1105..1152
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1390..1408)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1476..1580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1581..1649,1650..2441)
                     /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(1799..1818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2612..3200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3101..3120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3354..3371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3386..3525
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3451..3470)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3611..3633
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3627..3818)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5130..6212)
                     /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     6174..6193
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6213..6290)
                     /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 tcaccaagtt cttctatagc gatcagaatg
      421 tggatagccg tgatccggtg cagctgaatc tgctgtatgt tcaggcacgt gatgatattc
      481 tgaatggtag ccatagcgtg agctttgata aagcatgtga atttgcaggt tttcagtgcc
      541 agattcagtt tggtccgcat aatgaacaga aacacaaagc cggttttctg gatctgaaag
      601 attttctgcc gaaagaatac gtgaaacaga aaggtgaacg caaaattttc caggcgcata
      661 aaaactgtgg tcagatgagc gaaattgaag ccaaagttcg ttatgttaaa ctggcacgta
      721 gcctgaaaac atatggtgtt agcttttttc tggtgaaaga aaaaatgaaa ggcaaaaaca
      781 aactggttcc gcgtctgctg ggtattacca aagaatgtgt tatgcgtgtg gacgagaaaa
      841 ccaaagaagt tattcaagaa tggtccctga ccaacattaa acgttgggca gcaagcccga
      901 aaagctttac cctggatttt ggtgattatc aggatggcta ttattcagtt cagaccaccg
      961 aaggtgagca gattgcacag ctgattgcag gttatattga catcatcctg aaaaaaaaga
     1021 aaagctaaaa gggcgagctc agatccggct gctaacaaag cccgaaagga agctgagttg
     1081 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1141 aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta
     1201 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
     1261 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
     1321 taccgcatta aagctagctt atcgatgata agctgtcaaa catgagaatt aattcttgaa
     1381 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1441 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1501 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1561 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1621 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1681 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1741 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1801 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1861 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1921 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1981 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2041 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2101 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     2161 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2221 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2281 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2341 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2401 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2461 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3301 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3361 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3421 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3481 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3541 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3601 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3661 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3721 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3781 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3841 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3901 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     3961 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4021 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4081 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4141 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4201 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4261 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     4321 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     4381 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     4441 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     4501 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4561 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     4621 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4681 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4741 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4801 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4861 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4921 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4981 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     5041 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5101 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5161 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5221 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5281 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5341 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5401 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5461 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5521 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5581 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5641 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5701 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5761 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5821 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     5881 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     5941 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     6001 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     6061 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6121 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6181 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6241 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6301 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6361 gagcaccgcc gccgcaagga atggtgcatg
//
