LOCUS       sequence-325969-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human talin2 head.
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
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             13..36
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      35..44
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1252..1284
                     /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             1334..1351
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1404..1422)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1418..1465
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1502..1957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1589..1608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1799..1820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2049..2864)
                     /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(2049..2864)
                     /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     2772..2791
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2856..2875
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2986..3574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3475..3494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3728..3745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3760..3899
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3825..3844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3985..4007
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4001..4192)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5001..6083)
                     /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     6045..6064
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6084..6161)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6367..6386
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6470..6489
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6470..6488
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6489..6513
                     /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             6528..6550
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(6569..6580,1..12)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
ORIGIN
        1 catcaccatc atctggaagt tctgttccag gggcccatgg tggccctgtc cttaaagatt
       61 tgtgtgcgcc actgcaacgt ggtgaagacc atgcagtttg aaccatctac agctgtgtac
      121 gatgcgtgtc gagtcattcg ggaacgggtg cctgaggcac aagctgggca agcttctgac
      181 tatggactct ttctttcgga tgaagacccg aggaaaggga tttggctgga agcgggcaga
      241 acactggatt actacatgtt gcggaatggg gatattttgg aatataaaaa gaaacagaga
      301 cctcagaaaa tccggatgct ggatggatct gtgaagacag tgatggtgga tgattccaag
      361 actgtggggg agctcctggt cactatttgt agcagaatag gaataacaaa ttatgaagaa
      421 tactccttaa tccaagaaac tatcgaagaa aagaaagagg aaggaacggg cacactcaaa
      481 aaagacagga cactgttacg agatgagagg aaaatggaga agttgaaggc caagctgcac
      541 acagatgatg acctaaattg gctggatcac agccgaacat tcagagaaca aggagtagat
      601 gaaaacgaaa cgttgctgct tagacggaag ttcttttact ctgatcagaa tgtagattcg
      661 agagaccccg tgcagctgaa cttgctttat gttcaggcac aggatgacat cctgaatggc
      721 tctcaccctg tctccttcga gaaagcttgc gagtttggtg gatttcaagc ccagatacaa
      781 tttggacctc atgtggaaca taaacacaaa cctggatttt tagatctgaa ggaattcctg
      841 cccaaagaat acatcaagca gagaggagct gaaaagagga tctttcagga gcataagaac
      901 tgcggagaga tgagcgagat agaagccaag gtcaagtacg tcaaactcgc acggtccctc
      961 cgcacatatg gcgtgtcctt cttcctggtg aaggagaaga tgaaaggcaa gaacaagctg
     1021 gtgcctcgcc tgctggggat caccaaagac tcggtgatgc gcgtggatga gaagaccaag
     1081 gaagtgctgc aggagtggcc cctcaccacc gtcaagcgct gggcagcctc acccaagagc
     1141 ttcacactgg attttgggga gtatcaggaa agctactatt cagtacaaac caccgaggga
     1201 gagcagatat cccagctgat tgcaggctac attgacatca tcctgtgaca tatggctagc
     1261 atgactggtg gacagcaaat gggtcgcgga tccgaattcg agctccgtcg acaagcttgc
     1321 ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     1381 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1441 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     1501 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1561 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1621 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1681 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1741 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1801 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1861 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1921 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     1981 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2041 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     2101 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     2161 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     2221 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     2281 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     2341 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     2401 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     2461 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     2521 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     2581 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     2641 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     2701 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     2761 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     2821 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     2881 ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc
     2941 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3001 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3061 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3121 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3361 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3601 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3661 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3721 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     3781 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     3841 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     3901 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3961 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4021 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4081 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     4141 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     4201 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     4261 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     4321 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     4381 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     4441 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     4501 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     4561 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     4621 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     4681 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     4741 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     4801 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     4861 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     4921 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     4981 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5041 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     5101 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     5161 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     5221 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     5281 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     5341 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     5401 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     5461 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     5521 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     5581 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     5641 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     5701 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     5761 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     5821 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     5881 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     5941 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     6001 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     6061 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     6121 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     6181 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     6241 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     6301 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     6361 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     6421 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     6481 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     6541 aagaaggaga tataccatgg gcagcagcca tcatcatcat
//
