LOCUS       sequence-214612-        6009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expresses the unstructured neck region of mouse talin2 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6009)
  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 6009)
  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_Neck
FEATURES             Location/Qualifiers
     source          1..6009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             429..620
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(614..636)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    722..861
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     777..796
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(876..893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1047..1635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1127..1146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1806..2597,2598..2666))
                     /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     2429..2448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2667..2771)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2839..2857
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3095..3142)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3138..3156
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3486..3506)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3507..3548)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3549..3566)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3577..3599
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3614..3638
                     /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)."
     primer_bind     complement(3638..3657)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3639..3657)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3741..3760)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3966..4043
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4044..5126
                     /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     complement(4063..4082)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
       61 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
      121 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
      181 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
      241 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
      301 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
      361 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
      421 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
      481 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
      541 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
      601 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
      661 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
      721 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
      781 atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca cagatgcgta
      841 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
      901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     1501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2101 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2401 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     2461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2761 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2821 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattaatt
     2881 ctcatgtttg acagcttatc atcgataagc tagctttaat gcggtagttt atcacagtta
     2941 aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc
     3001 ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc
     3061 ttgcgggata tccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag
     3121 aggccccaag gggttatgct agttattgct cagcggtggc agcagccaac tcagcttcct
     3181 ttcgggcttt gttagcagcc ggatctgagc tcgccctttt aacgatgcat ctgaccaacc
     3241 ataacctgct gctgagggct cggcatgcta ccaacattaa aggtttccgg accgctgcta
     3301 ccgctacgca taactgcagg cagtgcaacg ctaccatgtt ctgctttacc ggtacgatta
     3361 aactgctgct gcagaatggt actttttttc gggctaacgc tttcttccag catggtgctt
     3421 tcttcatcac cttccagacc aaagcgatct ttgctctgtt tttttttcag ggtgaaggga
     3481 tcaattccct gaaaatacag gttttccgta gaatcgagac cgaggagagg gttagggata
     3541 ggcttaccat ggtgatggtg atgatgcata tgtatatctc cttcttaaag ttaaacaaaa
     3601 ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt
     3661 tcgcgggatc gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     3721 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     3781 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     3841 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     3901 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga
     3961 tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     4021 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     4081 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     4141 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     4201 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     4261 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     4321 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     4381 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     4441 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     4501 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     4561 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     4621 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     4681 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     4741 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     4801 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     4861 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     4921 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     4981 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     5041 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5101 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     5161 taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     5221 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     5281 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     5341 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     5401 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     5461 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc
     5521 cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg
     5581 tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc
     5641 agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc
     5701 acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg
     5761 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg
     5821 ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg
     5881 cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc
     5941 ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt
     6001 tgaggaccc
//
