LOCUS       sequence-330256-        5629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of human talin head (residues 1-400) 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 5629)
  AUTHORS   Zhang P, Azizi L, Kukkurainen S, Gao T, Baikoghli M, Jacquier MC, 
            Sun Y, Maatta JAE, Cheng RH, Wehrle-Haller B, Hytonen VP, Wu J
  TITLE     Crystal structure of the FERM-folded talin head reveals the 
            determinants for integrin binding.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Dec 22;117(51):32402-32412. doi: 
            10.1073/pnas.2014583117. Epub 2020 Dec 7.
  PUBMED    33288722
REFERENCE   2  (bases 1 to 5629)
  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: 
            "10.1073/pnas.2014583117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-12-22- 22"; volume: "117"; issue: "51"; pages: 
            "32402-32412"
COMMENT     Sequence Label: t1-400
FEATURES             Location/Qualifiers
     source          1..5629
                     /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(1752..1770)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1753..1770)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1813..1830)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1983..2069
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2161..2188
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2207..2298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2299..2367,2368..3159)
                     /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(2517..2536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3330..3918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3819..3838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4072..4089
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4104..4244
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4170..4189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4330..4352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4430..4507
                     /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             4508..5590
                     /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(4527..4546)
                     /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 aaggaggaaa taacagggac
      961 cttaagaaag gacaagacat tgctgcgaga tgaaaagaag atggagaaac taaagcagaa
     1021 attgcacaca gatgatgagt tgaactggct ggaccatggt cggacactga gggagcaggg
     1081 tgtagaggag cacgagacgc tgctgctgcg gaggaagttc ttttactcag accagaatgt
     1141 ggattcccgg gaccctgtac agctgaacct cctgtatgtg caggcacgag atgacatcct
     1201 gaatggctcc caccctgtct cctttgacaa ggcctgtgag tttgctggct tccaatgcca
     1261 gatccagttt gggccccaca atgagcagaa gcacaaggct ggcttccttg acctgaagga
     1321 cttcctgccc aaggagtatg tgaagcagaa gggagagcgt aagatcttcc aggcacacaa
     1381 gaattgtggg cagatgagtg agattgaggc caaggtccgc tacgtgaagc tagcccgttc
     1441 tctcaagact tacggtgtct ccttcttcct ggtgaaggaa aaaatgaaag ggaagaacaa
     1501 gctagtgccc aggcttctgg gcatcaccaa ggagtgtgtg atgcgagtgg atgagaagac
     1561 caaggaagtg atccaggagt ggaacctcac caacatcaaa cgctgggctg cgtctcccaa
     1621 aagcttcacc ctggattttg gagattacca agatggctat tactcagtac agacaactga
     1681 aggggagcag attgcacagc tcattgccgg ctacatcgat atcatcctgt aatgagtcta
     1741 gagggcccta ttctatagtg tcacctaaat gctaaagctt ggctgttttg gcggatgaga
     1801 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     1861 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     1921 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     1981 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     2041 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     2101 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     2161 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     2221 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2281 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2341 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2401 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2461 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2521 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     2581 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2641 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2701 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2761 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2821 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2881 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2941 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3001 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3061 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3121 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3181 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3241 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3781 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3841 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3901 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3961 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4021 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4081 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4141 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4201 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     4261 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4321 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4381 ggcagcagat caattcgcgc gcgaaggcga agcggcatgc atttacgttg acaccatcga
     4441 atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     4501 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     4561 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     4621 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     4681 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     4741 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     4801 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     4861 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     4921 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     4981 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     5041 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     5101 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     5161 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     5221 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     5281 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     5341 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     5401 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     5461 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     5521 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5581 cgggcagtga gcgcaacgca attaatgtaa gttagcgcga attgatctg
//
