LOCUS       sequence-330258-        5542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human talin-1 head (aa1-405) in E. coli. Contains 37 
            amino acid deletion in F1-loop, replaced with Gly-Ala-Gly. 
            N-terminal His6, Xpress-epitopeDLYDDDDK) and enterokinase cleavage 
            site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5542)
  AUTHORS   Kukkurainen S, Azizi L, Zhang P, Jacquier MC, Baikoghli M, von Essen
            M, Tuukkanen A, Laitaoja M, Liu X, Rahikainen R, Orlowski A, Janis 
            J, Maatta JAE, Varjosalo M, Vattulainen I, Rog T, Svergun D, Cheng 
            RH, Wu J, Hytonen VP, Wehrle-Haller B
  TITLE     The F1 loop of the talin head domain acts as a gatekeeper in 
            integrin activation and clustering.
  JOURNAL   J Cell Sci. 2020 Oct 12;133(19). pii: 133/19/jcs239202. doi: 
            10.1242/jcs.239202.
  PUBMED    33046605
REFERENCE   2  (bases 1 to 5542)
  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"; journalName: "J Cell 
            Sci."; date: "2020-10-12"; volume: "133(19). pii: 133/19/jcs239202. 
            doi"; pages: " 10.1242/jcs.239202"
COMMENT     Sequence Label: t1-405(del37/GAG)
FEATURES             Location/Qualifiers
     source          1..5542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     primer_bind     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             446..478
                     /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             join(482..490,491..505)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     promoter        complement(1665..1683)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1666..1683)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1726..1743)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1896..1982
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2074..2101
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2120..2211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2212..2280,2281..3072)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2430..2449)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3243..3831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3732..3751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3985..4002
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4017..4157
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4083..4102)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4243..4265
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4343..4420
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4421..5503
                     /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     complement(4440..4459)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gacgatgacg ataaggatcg atggatccga cctcgagcca tggttgcact
      541 ttcactgaag atcagcattg ggaatgtggt gaagacgatg cagtttgagc cgtctaccat
      601 ggtgtacgac gcctgccgca tcattcgtga gcggatccca gaggccccag ctggtcctcc
      661 cagcgacttt gggctctttc tgtcagatga tgaccccaaa aagggtatat ggctggaggc
      721 tgggaaagct ttggactact acatgctccg aaatggggac actatggagt acaggaagaa
      781 acagagaccc ctgaagatcc gtatgctgga tggaactgtg aagacgatca tggtggatga
      841 ctctaagact gtcactgaca tgctcatgac catctgtgcc cgcattggca tcaccaatca
      901 tgatgaatat tcattggttc gagagctggg cgcagggttg aactggctgg accatggtcg
      961 gacactgagg gagcagggtg tagaggagca cgagacgctg ctgctgcgga ggaagttctt
     1021 ttactcagac cagaatgtgg attcccggga ccctgtacag ctgaacctcc tgtatgtgca
     1081 ggcacgagat gacatcctga atggctccca ccctgtctcc tttgacaagg cctgtgagtt
     1141 tgctggcttc caatgccaga tccagtttgg gccccacaat gagcagaagc acaaggctgg
     1201 cttccttgac ctgaaggact tcctgcccaa ggagtatgtg aagcagaagg gagagcgtaa
     1261 gatcttccag gcacacaaga attgtgggca gatgagtgag attgaggcca aggtccgcta
     1321 cgtgaagcta gcccgttctc tcaagactta cggtgtctcc ttcttcctgg tgaaggaaaa
     1381 aatgaaaggg aagaacaagc tagtgcccag gcttctgggc atcaccaagg agtgtgtgat
     1441 gcgagtggat gagaagacca aggaagtgat ccaggagtgg aacctcacca acatcaaacg
     1501 ctgggctgcg tctcccaaaa gcttcaccct ggattttgga gattaccaag atggctatta
     1561 ctcagtacag acaactgaag gggagcagat tgcacagctc attgccggct acatcgatat
     1621 catcctgaag aagaaaaaaa gctaatgagt ctagagggcc ctattctata gtgtcaccta
     1681 aatgctaaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1741 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1801 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1861 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1921 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1981 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     2041 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     2101 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2161 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2221 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2281 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2341 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2401 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2461 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2521 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2581 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2641 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2701 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2761 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2821 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2881 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2941 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3001 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3061 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3121 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3181 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3241 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3301 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3361 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3421 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3481 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3541 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3601 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3661 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3721 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3781 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3841 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3901 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3961 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4021 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     4081 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     4141 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4201 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4261 agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg
     4321 cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc gcggtatggc
     4381 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     4441 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     4501 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     4561 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     4621 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     4681 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     4741 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     4801 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     4861 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     4921 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     4981 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     5041 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     5101 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     5161 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     5221 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     5281 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     5341 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     5401 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5461 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5521 taagttagcg cgaattgatc tg
//
