LOCUS       sequence-380513-        6912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a variant of human talin1 domains R12-DD, with a deletion 
            between K2375 and A2397.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6912)
  AUTHORS   Elefant N, Rouni G, Arapatzi C, Oz-Levi D, Sion-Sarid R, Edwards WJ,
            Ball NJ, Yanovsky-Dagan S, Cowell AR, Meiner V, Vainstein V, 
            Grammenoudi S, Lancet D, Goult BT, Harel T, Kostourou V
  TITLE     Talin1 dysfunction is genetically linked to systemic capillary leak 
            syndrome.
  JOURNAL   JCI Insight. 2024 Dec 20;9(24):e173664. doi: 
            10.1172/jci.insight.173664.
  PUBMED    39704176
REFERENCE   2  (bases 1 to 6912)
  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: "JCI 
            Insight."; date: "2024-12-20"; volume: "9(24):e173664. doi"; pages: 
            " 10.1172/jci.insight.173664"
COMMENT     Sequence Label: hTln1-R12DD-SCLS
FEATURES             Location/Qualifiers
     source          1..6912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     216..235
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        216..234
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    235..259
                     /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             274..296
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             307..324
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             325..366
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             367..387
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1620..1638)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1634..1681
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1919..1937)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2005..2109
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2110..2178,2179..2970)
                     /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(2328..2347)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3141..3729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3630..3649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3883..3900
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3915..4054
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3980..3999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4140..4162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4156..4347)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5659..6741)
                     /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     6703..6722
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6742..6819)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
       61 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
      121 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      181 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
      241 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
      301 catatgcatc atcaccatca ccatggtaag cctatcccta accctctcct cggtctcgat
      361 tctacggaaa acctgtattt tcagggaatt gatcccttca ccgaagcaac caaaggcacc
      421 cgtgcactgg aagccaccac cgaacatatt cgtcaagaac tggcagtttt ttgtagtccg
      481 gaaccgcctg caaaaaccag cacaccggaa gattttattc gtatgaccaa aggtattacc
      541 atggcaaccg caaaagcagt tgcagcaggt aatagctgcc gtcaagaaga tgttattgca
      601 accgccaatc tgagccgtcg tgcaattgcc gatatgctgc gtgcatgtaa agaagcagca
      661 tttcatccgg aagttgcacc ggatgttcgt ctgcgtgcac tgcattatgg tcgtgaatgt
      721 gcaaatggtt atctggaact gctggatcat gttctgctga ccctgcagaa accgaatccg
      781 gatctgaaac agcagctgac aggtcatagc aaacgtgttg caggtagcgt taccgaactg
      841 attcaggcag cagaagccat gaaaggcacc gaatgggttg atccggaaga tccgaccgtt
      901 attgcagaaa atgagctgct gggtgcagca gcagcaattg aagcagccgc aaaaaaactg
      961 gaacagctga aaccgcgtgc caaaccgaaa gaagccgacg aaagcctgaa ttttgaagaa
     1021 caaattctgg aagcagccaa aagtattgcc gcagcaacca gcgcactggt taaagcagca
     1081 agcgcagcac agcgtgaact ggttgcacag ggtaaagcag cccgtatggt tgcggcagcc
     1141 accaataatc tgtgtgaagc agcaaatgca gcagttcagg gtcatgcaag ccaagaaaaa
     1201 ctgattagca gcgcaaaaca ggttgccgca agcaccgcac agctgctggt tgcctgtaaa
     1261 gttaaagccg atcaggatag cgaagcaatg aaacgtctgc aggcagccgg taatgcagtt
     1321 aaacgtgcaa gcgataacct ggtgaaagcc gcacagaaag cagcagcctt tgaagatcaa
     1381 gaaaatgaaa ccgttgtggt gaaagaaaaa atggttggtg gtattgcaca gatcattgca
     1441 gcacaagaag agatgctgcg taaagaacgc gaactggaag aagcacgtaa aaaactggcc
     1501 cagattcgtc agcagcagta taaattcctg ccgagcgaac tgcgtgatga acattaaaag
     1561 ggcgagctca gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1621 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1681 gctgaaagga ggaactatat ccggatatcc cgcaagaggc ccggcagtac cggcataacc
     1741 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
     1801 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
     1861 agctagctta tcgatgataa gctgtcaaac atgagaatta attcttgaag acgaaagggc
     1921 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     1981 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2041 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2101 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2161 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2221 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2281 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2341 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2401 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2461 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2521 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2581 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2641 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2701 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2761 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2821 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2881 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2941 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3001 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3061 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3121 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3181 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3241 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3301 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3361 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3421 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3481 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3541 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3601 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3661 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3721 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3781 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3841 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3901 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3961 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     4021 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     4081 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4141 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     4201 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     4261 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     4321 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     4381 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     4441 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     4501 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     4561 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     4621 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     4681 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     4741 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     4801 catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga
     4861 gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg
     4921 caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct
     4981 tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca
     5041 aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca
     5101 taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc
     5161 gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac
     5221 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
     5281 cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg
     5341 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     5401 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     5461 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     5521 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     5581 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     5641 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5701 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     5761 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     5821 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     5881 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     5941 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     6001 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     6061 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     6121 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     6181 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     6241 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     6301 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     6361 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     6421 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     6481 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     6541 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     6601 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     6661 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     6721 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     6781 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     6841 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     6901 ccgcaaggaa tg
//
