LOCUS       sequence-325962-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human talin2 head mutant .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Rangarajan ES, Primi MC, Colgan LA, Chinthalapudi K, Yasuda R, Izard
            T
  TITLE     A distinct talin2 structure directs isoform specificity in cell 
            adhesion.
  JOURNAL   J Biol Chem. 2020 Sep 11;295(37):12885-12899. doi: 
            10.1074/jbc.RA119.010789. Epub 2020 Jun 30.
  PUBMED    32605925
REFERENCE   2  (bases 1 to 6580)
  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"; doi: 
            "10.1074/jbc.RA119.010789"; journalName: "J Biol Chem"; date: 
            "2020-09-11- 11"; volume: "295"; issue: "37"; pages: "12885-12899"
COMMENT     Sequence Label: pET28_hTLN2-head-LBD
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             81..104
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             105..128
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      127..136
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1344..1376
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1426..1443
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1496..1514)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1510..1557
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1594..2049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1681..1700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1891..1912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2141..2956)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2141..2956)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2864..2883
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2948..2967
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3078..3666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3567..3586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3820..3837
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3852..3991
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3917..3936)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4077..4099
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4093..4284)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5093..6175)
                     /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     6137..6156
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6176..6253)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6459..6478
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6562..6580,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6562..6580
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatataccat gggcagcagc catcatcatc atcatcacca tcatctggaa gttctgttcc
      121 aggggcccat ggtggccctg tccttaaaga tttgtgtgcg ccactgcaac gtggtgaaga
      181 ccatgcagtt tgaaccatct acagctgtgt acgatgcgtg tcgagtcatt cgggaacggg
      241 tgcctgaggc acaagctggg caagcttctg actatggact ctttctttcg gatgaagacc
      301 cgaggaaagg gatttggctg gaagcgggca gaacactgga ttactacatg ttgcggaatg
      361 gggatatttt ggaatataaa aagaaacaga gacctcagaa aatccggatg ctggatggat
      421 ctgtgaagac agtgatggtg gatgattcca agactgtggg ggagctcctg gtcactattt
      481 gtagcagaat aggaataaca aattatgaag aatactcctt aatccaagaa actatcgaag
      541 aaaagaaaga ggaaggaacg ggcacactca aaaaagacag gacactgtta cgagatgaga
      601 ggaaaatgga gaagttgaag gccaagctgc acacagatga tgacctaaat tggctggatc
      661 acagccgaac attcagagaa caaggagtag atgaaaacga aacgttgctg cttagacgga
      721 agttctttta ctctgatcag aatgtagatt cgagagaccc cgtgcagctg aacttgcttt
      781 atgttcaggc acaggatgac atcctgaatg gctctcaccc tgtctccttc gagaaagctt
      841 gcgagtttgg tggatttcaa gcccagatac aatttggacc tcatgtggaa cataaacaca
      901 aacctggatt tttagatctg aaggaattcc tgcccaaaga atacatccag cagagaggag
      961 ctgaaaagag gatctttcag gagcataaga actgcggaga gatgagcgag atagaagcca
     1021 aggtcaagta cgtcaaactc gcacggtccc tccgcacata tggcgtgtcc ttcttcctgg
     1081 tgcaggagaa gatgaaaggc aagaaccagc tggtgcctcg cctgctgggg atcaccaaag
     1141 actcggtgat gcgcgtggat cagcagacca aggaagtgct gcaggagtgg cccctcacca
     1201 ccgtcaagcg ctgggcagcc tcacccaaga gcttcacact ggattttggg gagtatcagg
     1261 aaagctacta ttcagtacaa accaccgagg gagagcagat atcccagctg attgcaggct
     1321 acattgacat catcctgtga catatggcta gcatgactgg tggacagcaa atgggtcgcg
     1381 gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca ccaccaccac
     1441 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1501 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     1561 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     1621 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1681 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1741 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1801 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1861 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     1921 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1981 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     2041 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2101 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
     2161 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     2221 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2281 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2341 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     2401 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     2461 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     2521 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     2581 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     2641 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     2701 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     2761 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     2821 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     2881 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     2941 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     3001 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3061 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3121 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3181 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3601 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3661 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3721 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3781 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3841 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3901 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     3961 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4021 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4081 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     4141 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     4201 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     4261 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     4321 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     4381 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     4441 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     4501 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     4561 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     4621 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     4681 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     4741 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     4801 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     4861 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     4921 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     4981 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     5041 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     5101 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5161 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     5221 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     5281 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     5341 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     5401 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     5461 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     5521 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     5581 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     5641 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     5701 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     5761 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     5821 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     5881 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     5941 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     6001 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     6061 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     6121 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     6181 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     6241 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     6301 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     6361 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     6421 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     6481 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     6541 gagatctcga tcccgcgaaa ttaatacgac tcactatagg
//
