LOCUS       Exported                6153 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Plasmid expresses the talin2 rod sub domain R4 a 4 helix bundle..
ACCESSION   .
VERSION     .
KEYWORDS    mTalin2_R4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6153)
  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 6153)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6153
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     89..108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        89..107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    108..132
                     /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             147..169
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             180..197
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             198..239
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             240..260
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(734..752)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      748..795
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1033..1051)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1119..1223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1224..2084
                     /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(1442..1461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2255..2843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2744..2763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2997..3014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3029..3168
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3094..3113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3254..3276
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3270..3461)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4773..5855)
                     /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     5817..5836
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5856..5933)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6139..5
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
       61 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      121 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgc
      181 atcatcacca tcaccatggt aagcctatcc ctaaccctct cctcggtctc gattctacgg
      241 aaaacctgta ttttcaggga attgatccct tcaccgttaa tcgtctggaa gttgcagcaa
      301 aacaggcagc agcagccgca acccagacca ttgcagcaag ccagaatgca gcaattagca
      361 ataaaaaccc gagcgcacag cagcaactgg ttcagagctg taaagcagtt gcagatcata
      421 ttccgcagct ggttcagggt gttcgtggta gccaggcaca ggcagaagat ctgtcagcac
      481 agctggcact gattattagc agccagaatt ttctgcagcc tggtagcaaa atggttagca
      541 gcgcaaaagc agcagttccg accgttagcg atcaggcagc ggcaatgcag ctgagccagt
      601 gtgcaaaaaa tctggcaacc agcctggcag aactgcgtac cgccagccag aaagcacatg
      661 aagcatgtta aaagggcgag ctcagatccg gctgctaaca aagcccgaaa ggaagctgag
      721 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      781 ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca
      841 gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga
      901 tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata
      961 aactaccgca ttaaagctag cttatcgatg ataagctgtc aaacatgaga attaattctt
     1021 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1081 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1141 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1201 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1261 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1321 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1381 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1441 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     1501 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1561 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1621 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1681 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1741 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     1801 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1861 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     1921 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     1981 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2041 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2101 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2161 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2221 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2281 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2341 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2401 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2461 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2521 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2581 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2641 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2701 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2761 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2821 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2881 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2941 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3001 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3061 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3121 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3181 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3241 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3301 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3361 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3421 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3481 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     3541 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     3601 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     3661 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     3721 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     3781 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     3841 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     3901 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc
     3961 gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca
     4021 ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg
     4081 aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg
     4141 cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc
     4201 tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg
     4261 taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca
     4321 gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga
     4381 aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     4441 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     4501 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     4561 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     4621 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     4681 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     4741 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4801 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4861 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4921 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4981 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     5041 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     5101 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5161 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     5221 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     5281 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     5341 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     5401 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     5461 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     5521 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     5581 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     5641 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     5701 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     5761 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     5821 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     5881 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     5941 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     6001 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     6061 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6121 agcccgatct tccccatcgg tgatgtcggc gat
//
