LOCUS       sequence-330268-        5029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of human talin-1 head fragment (residues 206-405) in 
            E. coli. 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 5029)
  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 5029)
  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: t206-405
FEATURES             Location/Qualifiers
     source          1..5029
                     /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(1152..1170)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1153..1170)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1213..1230)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1383..1469
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1561..1588
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1607..1698
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1699..1767,1768..2559)
                     /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(1917..1936)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2730..3318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3219..3238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3472..3489
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3504..3644
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3570..3589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3730..3752
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3830..3907
                     /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             3908..4990
                     /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(3927..3946)
                     /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 cctcgagctt cccgggaccc
      541 tgtacagctg aacctcctgt atgtgcaggc acgagatgac atcctgaatg gctcccaccc
      601 tgtctccttt gacaaggcct gtgagtttgc tggcttccaa tgccagatcc agtttgggcc
      661 ccacaatgag cagaagcaca aggctggctt ccttgacctg aaggacttcc tgcccaagga
      721 gtatgtgaag cagaagggag agcgtaagat cttccaggca cacaagaatt gtgggcagat
      781 gagtgagatt gaggccaagg tccgctacgt gaagctagcc cgttctctca agacttacgg
      841 tgtctccttc ttcctggtga aggaaaaaat gaaagggaag aacaagctag tgcccaggct
      901 tctgggcatc accaaggagt gtgtgatgcg agtggatgag aagaccaagg aagtgatcca
      961 ggagtggaac ctcaccaaca tcaaacgctg ggctgcgtct cccaaaagct tcaccctgga
     1021 ttttggagat taccaagatg gctattactc agtacagaca actgaagggg agcagattgc
     1081 acagctcatt gccggctaca tcgatatcat cctgaagaag aaaaaaagct aatgagtcta
     1141 gagggcccta ttctatagtg tcacctaaat gctaaagctt ggctgttttg gcggatgaga
     1201 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     1261 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     1321 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     1381 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     1441 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     1501 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     1561 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     1621 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1681 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1741 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1801 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1861 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1921 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1981 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2041 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2101 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2161 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2221 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2281 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2341 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2401 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2461 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2521 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2581 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2641 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2701 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2761 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2821 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2881 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2941 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3001 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3061 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3121 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3181 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3241 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3301 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3361 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3421 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3481 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3541 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3601 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3661 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3721 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3781 ggcagcagat caattcgcgc gcgaaggcga agcggcatgc atttacgttg acaccatcga
     3841 atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     3901 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     3961 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     4021 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     4081 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     4141 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     4201 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     4261 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     4321 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     4381 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     4441 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     4501 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     4561 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     4621 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     4681 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     4741 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     4801 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     4861 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     4921 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4981 cgggcagtga gcgcaacgca attaatgtaa gttagcgcga attgatctg
//
