LOCUS       Exported                5858 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Recombinant expression of RBMS2 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    RBMS2_2xRRM_pGEX
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  AUTHORS   Dominguez D, Freese P, Alexis MS, Su A, Hochman M, Palden T, Bazile 
            C, Lambert NJ, Van Nostrand EL, Pratt GA, Yeo GW, Graveley BR, Burge
            CB
  TITLE     Sequence, Structure, and Context Preferences of Human RNA Binding 
            Proteins.
  JOURNAL   Mol Cell. 2018 Jun 7;70(5):854-867.e9. doi: 
            10.1016/j.molcel.2018.05.001. Epub 2018 Jun 7.
  PUBMED    29883606
REFERENCE   2  (bases 1 to 5858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5858
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        535..565
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    573..589
                     /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     597..613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     597..613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             609..1136
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(629..646)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(629..645)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(638..660)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1238..1266
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1274..1290
                     /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)."
     CDS             1313..1966
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1924..1946
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1973..1996
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2006..2119
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     primer_bind     complement(2963..2985)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3130..3148)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3216..3320
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3321..4181
                     /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(3539..3558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4352..4940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4841..4860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5094..5111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5184..5261
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5262..486
                     /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(5281..5300)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
       61 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
      121 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
      181 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
      241 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
      301 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
      361 cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca
      541 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      601 aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg
      661 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      721 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      781 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
      841 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
      901 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg
      961 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     1021 gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca cggtgcacca
     1081 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca
     1141 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac
     1201 atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt
     1261 ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagtat tcatgtcccc
     1321 tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata
     1381 tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa
     1441 caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt
     1501 taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg
     1561 tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag
     1621 atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct
     1681 tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt
     1741 aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt
     1801 atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat
     1861 tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt
     1921 gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt
     1981 tctgttccag gggcccctgg gattaatgga cgaaaaaacc accggttggc gtggtggtca
     2041 cgttgttgaa ggtctggctg gtgaactgga acagctgcgt gctcgtctgg aacaccaccc
     2101 gcagggtcag cgtgaaccgg gatccatgct gctatccgtg acttccaggc ccgggatttc
     2161 gacttttggc tacaatagaa acaacaagaa gccatatgtg tcactggctc agcagatggc
     2221 accacctagc ccaagcaaca gtacacctaa cagcagtagt ggaagcaatg gaaatgacca
     2281 gctgagcaaa accaacctat acatccgagg attgcaacca ggcactactg accaagatct
     2341 tgtcaagctg tgtcagccat atggcaagat tgtttccact aaggccatac tggacaagac
     2401 cacaaacaaa tgtaaaggct atggctttgt agattttgac agcccttcag cagcacagaa
     2461 agctgtaaca gcactgaagg ccagcggtgt acaggcacag atggcaaagc aacaggaaca
     2521 ggaccccaca aatttataca tctcaaacct cccactgtca atggatgagc aggaactgga
     2581 ggggatgctg aagccctttg gccaggttat ctccacccgt atccttcgag ataccagtgg
     2641 gaccagcaga ggtgttggct ttgcaaggat ggagtccaca gagaagtgtg aagccatcat
     2701 cacccacttt aatggaaaat atattaagac accccctgga gtaccagccc catccgatcc
     2761 cttgctttgc aaatttgctg atggcgggcc aaagaaacga cagaaccaag gaaaatttgt
     2821 gcaaaatgga cgggcttggc caaggaatgc agacatgggc gtcatggcct tgacctatga
     2881 ccccaccaca gctcttcagt gagcggccgc atcgtgactg actgacgatc tgcctcgcgc
     2941 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3001 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3061 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa
     3121 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3181 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3241 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3301 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3361 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3421 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3481 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3541 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     3601 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3661 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3721 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3781 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3841 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     3901 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3961 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4021 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4081 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4141 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4201 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4261 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4321 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4381 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4441 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4501 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4561 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4621 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4681 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4741 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4801 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4861 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4921 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4981 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5041 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5101 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5161 gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg
     5221 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     5281 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     5341 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     5401 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     5461 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     5521 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     5581 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     5641 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     5701 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     5761 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     5821 gcccattaag ttctgtctcg gcgcgtctgc gtctggct
//
