LOCUS       Exported                5679 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression of the SH3 domain of human Fyn.
ACCESSION   .
VERSION     .
KEYWORDS    Fyn-SH3_5WT_S25C
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5679)
  AUTHORS   Amaya J, Ryan VH, Fawzi NL
  TITLE     The SH3 domain of Fyn kinase interacts with and induces 
            liquid-liquid phase separation of the low-complexity domain of 
            hnRNPA2.
  JOURNAL   J Biol Chem. 2018 Dec 21;293(51):19522-19531. doi: 
            10.1074/jbc.RA118.005120. Epub 2018 Nov 5.
  PUBMED    30397184
REFERENCE   2  (bases 1 to 5679)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5679
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             98..115
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             146..166
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             411..428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(481..499)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      495..542
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      579..1034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(666..685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     876..897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1060..1164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1165..2025
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1383..1402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2196..2784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2685..2704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2938..2955
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2970..3109
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3035..3054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3195..3217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3211..3402)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4211..5293)
                     /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     5255..5274
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5294..5371)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5577..5596
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gatgaaacac caccaccacc accacccgat
      121 gagcgactac gatattccga cgaccgaaaa tctgtacttc caaggcgcaa tggttcaaat
      181 tagtaccctg tttgaagcgc tgtatgatta cgaagcccgt acggaagatg acctgtgctt
      241 tcataaaggc gaaaaattcc agatcctgaa cagctctgag ggtgactggt gggaagcgcg
      301 cagcctgacg acgggtgaaa cgggctacat tccgagtaac tacgtggctc cggtggatcg
      361 ctgaggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag caccaccacc
      421 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
      481 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      541 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
      601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
      661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
      721 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      781 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      841 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      901 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
      961 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1021 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1081 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1141 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1201 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1261 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1321 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1381 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1441 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1501 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1561 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1621 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1681 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     1741 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1801 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1861 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1921 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1981 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2041 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2101 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2161 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2221 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2281 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2341 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2401 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2461 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2521 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2581 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2641 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2701 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2761 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2821 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2881 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2941 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     3001 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3061 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     3121 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3181 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3241 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     3301 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     3361 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     3421 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     3481 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     3541 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     3601 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     3661 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     3721 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     3781 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     3841 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt
     3901 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     3961 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     4021 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     4081 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     4141 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     4201 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4261 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     4321 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     4381 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     4441 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     4501 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     4561 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     4621 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     4681 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     4741 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     4801 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4861 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4921 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4981 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     5041 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     5101 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     5161 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     5221 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     5281 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     5341 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     5401 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     5461 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5521 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5581 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5641 cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
