LOCUS       sequence-213042-        6234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the talin2 rod domain R12 a 5 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6234)
  AUTHORS   Gough RE, Goult BT
  TITLE     The tale of two talins - two isoforms to fine-tune integrin 
            signalling.
  JOURNAL   FEBS Lett. 2018 May 3. doi: 10.1002/1873-3468.13081.
  PUBMED    29723415
REFERENCE   2  (bases 1 to 6234)
  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: "FEBS Lett. 
            2018 May 3. doi: 10.1002/1873-3468.13081."
COMMENT     Sequence Label: mTalin2_R12
FEATURES             Location/Qualifiers
     source          1..6234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        58..76
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    77..101
                     /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             116..138
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..166
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             167..208
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             209..229
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(784..802)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      798..845
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1083..1101)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1169..1273
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1274..1342,1343..2134)
                     /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(1492..1511)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2305..2893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2794..2813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3047..3064
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3079..3218
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3144..3163)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3304..3326
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3320..3511)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4823..5905)
                     /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     5867..5886
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5906..5983)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6189..6208
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
       61 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
      121 ttaactttaa gaaggagata tacatatgca tcatcaccat caccatggta agcctatccc
      181 taaccctctc ctcggtctcg attctacgga aaacctgtat tttcagggaa ttgatccctt
      241 caccgatgaa gcaacccgtg gcacccgtgc actggaagca accattgaat atatcaaaca
      301 agaactgacc gtgtttcaga gcaaagatat tccggaaaaa accagcagtc cggaagaaag
      361 cattcgtatg accaaaggta ttaccatggc aaccgcaaaa gcagttgcag caggtaatag
      421 ctgtcgtcaa gaagatgtta ttgcaaccgc caatctgagc cgtaaagcag ttagcgatat
      481 gctgattgca tgtaaacagg caagctttta tccggatgtt agcgaagaag ttcgtacacg
      541 cgcactgcgt tatggcaccg aatgtaccct gggttatctg gatctgctgg aacatgtgct
      601 ggttatcctg cagaaaccga caccggaact gaaacatcag ctggcagcat ttagcaaacg
      661 tgttgccggt gcagttaccg aactgattca ggcagcagaa gcaatgaaag gtacagaata
      721 aaagggcgag ctcagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      781 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      841 ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat
      901 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
      961 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     1021 ttaaagctag cttatcgatg ataagctgtc aaacatgaga attaattctt gaagacgaaa
     1081 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1141 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1201 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1261 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1321 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1381 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1441 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1501 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     1561 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1621 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1681 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1741 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1801 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1861 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1921 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1981 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2041 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2101 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2161 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2221 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2521 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2581 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2641 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2701 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2761 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2821 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2881 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2941 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3001 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3061 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3121 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3181 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3241 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3301 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3361 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     3421 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3481 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     3541 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     3601 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     3661 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     3721 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     3781 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     3841 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     3901 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     3961 cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc
     4021 tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc
     4081 tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc
     4141 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
     4201 gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc
     4261 ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc
     4321 gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     4381 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     4441 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
     4501 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     4561 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     4621 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     4681 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     4741 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     4801 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4861 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     4921 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     4981 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     5041 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     5101 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     5161 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     5221 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     5281 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     5341 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     5401 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     5461 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     5521 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     5581 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     5641 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     5701 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     5761 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     5821 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     5881 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     5941 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     6001 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     6061 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     6121 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     6181 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccg
//
