LOCUS       sequence-313952-        6147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human TLNRD1 4-helix domain (residues 143-273) in 
            bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6147)
  AUTHORS   Cowell AR, Jacquemet G, Singh AK, Varela L, Nylund AS, Ammon YC, 
            Brown DG, Akhmanova A, Ivaska J, Goult BT
  TITLE     Talin rod domain-containing protein 1 (TLNRD1) is a novel 
            actin-bundling protein which promotes filopodia formation.
  JOURNAL   J Cell Biol. 2021 Sep 6;220(9). pii: 212472. doi: 
            10.1083/jcb.202005214. Epub 2021 Jul 15.
  PUBMED    34264272
REFERENCE   2  (bases 1 to 6147)
  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 
            Biol."; date: "2021-09-6"; volume: "220(9). pii: 212472. doi"; 
            pages: " 10.1083/jcb.202005214. Epub 2021 Jul 15"
COMMENT     Sequence Label: human TLNRD1-4H pET151
FEATURES             Location/Qualifiers
     source          1..6147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..151
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             152..172
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(640..658)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      654..701
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(939..957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1025..1129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1130..1198,1199..1990)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1348..1367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2161..2749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2650..2669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2903..2920
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2935..3074
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3000..3019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3160..3182
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3176..3367)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4679..5761)
                     /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     5723..5742
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5762..5839)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6045..6064
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcatcatcac catcaccatg gtaagcctat
      121 ccctaaccct ctcctcggtc tcgattctac ggaaaacctg tattttcagg gaggatccgc
      181 acagcctggt ctggttgatc gttatcgtgt tacccgttgt cgtcatgaag ttgaacaggg
      241 ttgtgcagtt ctgcgtgcca caccgctggc agatatgaca ccgcagctgc tgttagaagt
      301 tagccagggt ctgagccgta atctgaaatt tctgaccgat gcatgtgcac tggcaagcga
      361 taaaagccgt gatcgtttta gccgtgaaca gttcaaactg ggtgttaaat gtatgagcac
      421 cagcgcaagc gcactgctgg catgtgttcg tgaagttaaa gttgcaccga gcgaactggc
      481 acgtagccgt tgtgcactgt ttagcggtcc gctggttcag gcagttagcg cactggttgg
      541 ttttgcaacc gaaccgcagt ttctgggtcg ttaactcgag taagagctca gatccggctg
      601 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
      661 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
      721 ccggatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc
      781 cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg
      841 actgcgttag caatttaact gtgataaact accgcattaa agctagctta tcgatgataa
      901 gctgtcaaac atgagaatta attcttgaag acgaaagggc ctcgtgatac gcctattttt
      961 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1021 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1081 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1141 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1201 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1261 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1321 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     1381 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1441 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1501 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1561 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1621 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     1681 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1741 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1801 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1861 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1921 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1981 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2041 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2101 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2281 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2821 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2881 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     2941 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     3001 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3061 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3121 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3181 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3241 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3301 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3361 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     3421 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     3481 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     3541 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     3601 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     3661 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     3721 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     3781 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     3841 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
     3901 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     3961 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     4021 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     4081 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     4141 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     4201 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     4261 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     4321 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4381 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4441 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4501 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4561 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4621 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4681 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4741 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4801 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4861 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4921 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     4981 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5041 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     5101 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     5161 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5221 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5281 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5341 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5401 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5461 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5521 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5581 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5641 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5701 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5761 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5821 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     5881 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     5941 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     6001 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     6061 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     6121 aggatcgaga tctcgatccc gcgaaat
//
