LOCUS       sequence-380511-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a variant of the human TLN1 R1R2 domains containing a 17AA
            insert into R2 (exon 17b) in bacteria .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6738)
  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 6738)
  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-R1R2-17b
FEATURES             Location/Qualifiers
     source          1..6738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..124
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     208..227
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        208..226
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    227..251
                     /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             266..288
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             299..316
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             317..358
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             359..379
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1438..1456)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1452..1499
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1737..1755)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1823..1927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1928..1996,1997..2788)
                     /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(2146..2165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2959..3547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3448..3467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3701..3718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3733..3872
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3798..3817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3958..3980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3974..4165)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5477..6559)
                     /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     6521..6540
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6560..6637)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
       61 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      121 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      181 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      241 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgca
      301 tcatcaccat caccatggta agcctatccc taaccctctc ctcggtctcg attctacgga
      361 aaacctgtat tttcagggaa ttgatccctt cacccgtggt catatgcctc cgctgaccag
      421 cgcacagcag gcactgaccg gcaccattaa tagcagcatg caggcagttc aggcagcaca
      481 ggcaaccctg gatgattttg ataccctgcc accgctgggt caagatgcag caagcaaagc
      541 atggcgtaaa aacaaaatgg atgaaagcaa acacgaaatc catagccagg ttgatgcaat
      601 taccgcaggc accgcaagcg ttgttaatct gaccgcaggc gatccggcag aaaccgatta
      661 caccgcagtt ggttgtgcag ttaccaccat tagcagcaat ctgacagaaa tgagccgtgg
      721 tgttaaactg ctggcagcac tgctggaaga tgaaggtggt agcggtcgtc cgctgctgca
      781 ggcagcaaaa ggtctggcag gcgcagttag cgaactgctg cgtagtgcac agcctgcaag
      841 cgcagaaccg cgtcagaatc tgctgcaagc cgcaggtaat gttggtcagg caagcggtga
      901 actgttacag cagattggtg aaagcgatac cgatccgcat tttcagattt gtgcaagtcg
      961 tggtgcgggt gttcgtagcc ctccggatag cccgaccgat gcactgatgc agctggcaaa
     1021 agcagttgcg agcgcagcag cagccctggt tctgaaagcc aaaagcgttg cacagcgtac
     1081 cgaagatagt ggcctgcaga cccaggttat tgcagcagca acccagtgtg cactgagcac
     1141 cagccagctg gttgcatgta ccaaagttgt tgcaccgacc attagctcac cggtttgtca
     1201 agaacagctg gtagaagcag gtcgtctggt tgcaaaagcc gttgaaggtt gtgttagcgc
     1261 aagccaggca gccaccgaag atggtcagct gctgcgtggt gttggtgcag cagccacagc
     1321 agttacccag gcgctgaatg aattactgca gcatgttaaa gcacatgcca cctaaaaggg
     1381 cgagctcaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     1441 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1501 tgaaaggagg aactatatcc ggatatcccg caagaggccc ggcagtaccg gcataaccaa
     1561 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     1621 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     1681 ctagcttatc gatgataagc tgtcaaacat gagaattaat tcttgaagac gaaagggcct
     1741 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1801 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1861 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1921 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1981 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2041 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2101 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2161 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2221 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2281 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2341 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2401 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2461 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2521 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2581 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2641 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2701 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2821 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2881 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2941 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3001 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3061 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3781 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     3841 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3901 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3961 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     4021 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     4081 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4141 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     4201 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     4261 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     4321 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     4381 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     4441 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     4501 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     4561 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     4621 tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga
     4681 tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca
     4741 agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc
     4801 cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag
     4861 gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata
     4921 aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga
     4981 tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc
     5041 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg
     5101 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc
     5161 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     5221 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     5281 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     5341 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     5401 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     5461 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5521 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     5581 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     5641 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     5701 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     5761 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     5821 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5881 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5941 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     6001 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     6061 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     6121 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     6181 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     6241 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     6301 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     6361 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     6421 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     6481 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     6541 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     6601 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     6661 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     6721 gcaaggaatg gtgcatgc
//
