LOCUS       sequence-212968-        6111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses mouse talin2 FERM domain F1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6111)
  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 6111)
  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_F1
FEATURES             Location/Qualifiers
     source          1..6111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        96..114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    115..139
                     /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             154..176
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             187..204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             205..246
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             247..267
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(699..717)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      713..760
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(998..1016)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1084..1188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1189..1257,1258..2049)
                     /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(1407..1426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2220..2808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2709..2728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2962..2979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2994..3133
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3059..3078)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3219..3241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3235..3426)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4738..5820)
                     /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     5782..5801
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5821..5898)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(6104..6111,1..12)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
       61 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
      121 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
      181 catatgcatc atcaccatca ccatggtaag cctatcccta accctctcct cggtctcgat
      241 tctacggaaa acctgtattt tcagggaatt gatcccttca cccgtccgca gaaaattcgt
      301 atgctggatg gtagcgttaa aaccgttatg gttgatgata gcaaaaccgt tggtgaactg
      361 ctggttacca tttgtagccg tattggcatt accaactatg aagaatatag cctgatccaa
      421 gaaaccatcg aagaaaaaaa agaagaaggc accggcaccc tgaaaaaaga tcgtaccctg
      481 ctgcgtgatg aacgcaaaat ggaaaaactg aaagcaaaac tgcacaccga tgatgatctg
      541 aattggctgg atcatagccg tacctttcgt gaacagggtg ttgatgaaaa tgaaacactg
      601 ctgctgcgtc gcaaattttt ctatagcgat cagtaaaagg gcgagctcag atccggctgc
      661 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
      721 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
      781 cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
      841 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
      901 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gctagcttat cgatgataag
      961 ctgtcaaaca tgagaattaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     1021 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     1081 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1141 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1201 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1261 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1321 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1381 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     1441 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1501 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1561 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1621 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1681 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     1741 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1801 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1861 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1921 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1981 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2041 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2101 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2341 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     3061 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     3121 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3241 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3301 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3361 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3421 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     3481 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3541 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3601 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     3661 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3721 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3781 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3841 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     3901 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     3961 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     4021 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     4081 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     4141 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     4201 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     4261 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     4321 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     4381 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     4441 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     4501 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     4561 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     4621 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     4681 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     4741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4801 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     4861 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     4921 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     4981 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     5041 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     5101 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     5161 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     5221 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     5281 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     5341 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     5401 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     5461 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     5521 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     5581 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     5641 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     5701 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     5761 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     5821 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     5881 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     5941 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     6001 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     6061 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg t
//
