LOCUS       sequence-380512-        6687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the human TLN1 R1R2 domains in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6687)
  AUTHORS   Gallego-Paez LM, Edwards WJS, Chanduri M, Guo Y, Koorman T, Lee CY, 
            Grexa N, Derksen P, Yan J, Schwartz MA, Mauer J, Goult BT
  TITLE     TLN1 contains a cancer-associated cassette exon that alters talin-1 
            mechanosensitivity.
  JOURNAL   J Cell Biol. 2023 May 1;222(5):e202209010. doi: 
            10.1083/jcb.202209010. Epub 2023 Mar 6.
  PUBMED    36880935
REFERENCE   2  (bases 1 to 6687)
  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: "2023-05-1"; volume: "222(5):e202209010. doi"; pages: 
            " 10.1083/jcb.202209010. Epub 2023 Mar 6"
COMMENT     Sequence Label: hTln1-R1R2
FEATURES             Location/Qualifiers
     source          1..6687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     219..238
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        219..237
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    238..262
                     /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             277..299
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             310..327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             328..369
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             370..390
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1398..1416)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1412..1459
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1697..1715)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1783..1887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1888..1956,1957..2748)
                     /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(2106..2125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2919..3507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3408..3427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3661..3678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3693..3832
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3758..3777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3918..3940
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3934..4125)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5437..6519)
                     /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     6481..6500
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6520..6597)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
       61 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
      121 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
      181 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga
      241 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      301 atacatatgc atcatcacca tcaccatggt aagcctatcc ctaaccctct cctcggtctc
      361 gattctacgg aaaacctgta ttttcaggga attgatccct tcacccgtgg tcatatgcct
      421 ccgctgacca gcgcacagca ggcactgacc ggcaccatta atagcagcat gcaggcagtt
      481 caggcagcac aggcaaccct ggatgatttt gataccctgc caccgctggg tcaagatgca
      541 gcaagcaaag catggcgtaa aaacaaaatg gatgaaagca aacacgaaat ccatagccag
      601 gttgatgcaa ttaccgcagg caccgcaagc gttgttaatc tgaccgcagg cgatccggca
      661 gaaaccgatt acaccgcagt tggttgtgca gttaccacca ttagcagcaa tctgacagaa
      721 atgagccgtg gtgttaaact gctggcagca ctgctggaag atgaaggtgg tagcggtcgt
      781 ccgctgctgc aggcagcaaa aggtctggca ggcgcagtta gcgaactgct gcgtagtgca
      841 cagcctgcaa gcgcagaacc gcgtcagaat ctgctgcaag ccgcaggtaa tgttggtcag
      901 gcaagcggtg aactgttaca gcagattggt gaaagcgata ccgatccgca ttttcaggat
      961 gcactgatgc agctggcaaa agcagttgcg agcgcagcag cagccctggt tctgaaagcc
     1021 aaaagcgttg cacagcgtac cgaagatagt ggcctgcaga cccaggttat tgcagcagca
     1081 acccagtgtg cactgagcac cagccagctg gttgcatgta ccaaagttgt tgcaccgacc
     1141 attagctcac cggtttgtca agaacagctg gtagaagcag gtcgtctggt tgcaaaagcc
     1201 gttgaaggtt gtgttagcgc aagccaggca gccaccgaag atggtcagct gctgcgtggt
     1261 gttggtgcag cagccacagc agttacccag gcgctgaatg aattactgca gcatgttaaa
     1321 gcacatgcca cctaaaaggg cgagctcaga tccggctgct aacaaagccc gaaaggaagc
     1381 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1441 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatcccg caagaggccc
     1501 ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt gccgaggatg
     1561 acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca atttaactgt
     1621 gataaactac cgcattaaag ctagcttatc gatgataagc tgtcaaacat gagaattaat
     1681 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     1741 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1861 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2101 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2401 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     2461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2761 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2821 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3061 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3601 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3661 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3721 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     3781 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     3841 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3901 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3961 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     4021 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4081 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     4141 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     4201 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     4261 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     4321 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     4381 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     4441 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     4501 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     4561 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg
     4621 ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg
     4681 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
     4741 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     4801 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     4861 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
     4921 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     4981 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     5041 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
     5101 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     5161 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     5221 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     5281 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     5341 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     5401 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5521 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5581 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5641 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5701 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5761 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5821 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5881 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5941 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     6001 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     6061 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     6121 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     6181 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     6241 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6301 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6361 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6421 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6481 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6541 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6601 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6661 ggccgttgag caccgccgcc gcaagga
//
