LOCUS       sequence-330260-        5554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human talin-1 head (residues 1-405) in E. coli. 
            Contains 30 amino acid deletion in F1-loop. N-terminal His6-tag, 
            Xpress-epitope (DLYDDDDK) and enterokinase cleavage site for tag 
            removal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  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 5554)
  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(del30)
FEATURES             Location/Qualifiers
     source          1..5554
                     /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(1677..1695)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1678..1695)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1738..1755)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1908..1994
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2086..2113
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2132..2223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2224..2292,2293..3084)
                     /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(2442..2461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3255..3843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3744..3763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3997..4014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4029..4169
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4095..4114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4255..4277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4355..4432
                     /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             4433..5515
                     /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(4452..4471)
                     /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 gagagctgat ggaagagaaa aaggatgagt tgaactggct
      961 ggaccatggt cggacactga gggagcaggg tgtagaggag cacgagacgc tgctgctgcg
     1021 gaggaagttc ttttactcag accagaatgt ggattcccgg gaccctgtac agctgaacct
     1081 cctgtatgtg caggcacgag atgacatcct gaatggctcc caccctgtct cctttgacaa
     1141 ggcctgtgag tttgctggct tccaatgcca gatccagttt gggccccaca atgagcagaa
     1201 gcacaaggct ggcttccttg acctgaagga cttcctgccc aaggagtatg tgaagcagaa
     1261 gggagagcgt aagatcttcc aggcacacaa gaattgtggg cagatgagtg agattgaggc
     1321 caaggtccgc tacgtgaagc tagcccgttc tctcaagact tacggtgtct ccttcttcct
     1381 ggtgaaggaa aaaatgaaag ggaagaacaa gctagtgccc aggcttctgg gcatcaccaa
     1441 ggagtgtgtg atgcgagtgg atgagaagac caaggaagtg atccaggagt ggaacctcac
     1501 caacatcaaa cgctgggctg cgtctcccaa aagcttcacc ctggattttg gagattacca
     1561 agatggctat tactcagtac agacaactga aggggagcag attgcacagc tcattgccgg
     1621 ctacatcgat atcatcctga agaagaaaaa aagctaatga gtctagaggg ccctattcta
     1681 tagtgtcacc taaatgctaa agcttggctg ttttggcgga tgagagaaga ttttcagcct
     1741 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     1801 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     1861 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     1921 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     1981 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     2041 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     2101 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     2161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2221 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2281 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2341 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2401 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2461 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2521 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2581 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2641 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2701 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2761 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2821 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2881 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2941 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3001 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3061 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3121 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3361 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3901 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3961 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4021 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4081 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     4141 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     4201 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4261 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt
     4321 cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggt gcaaaacctt
     4381 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     4441 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     4501 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     4561 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     4621 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     4681 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     4741 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     4801 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     4861 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     4921 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     4981 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     5041 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     5101 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     5161 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     5221 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     5281 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     5341 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     5401 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5461 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5521 acgcaattaa tgtaagttag cgcgaattga tctg
//
