LOCUS       sequence-274001-        6671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of IGF2BP1 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  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 6671)
  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"; doi: 
            "10.1016/j.molcel.2018.05.001"; journalName: "Mol Cell"; date: 
            "2018-06-7- 7"; volume: "70"; issue: "5"; pages: "854-867.e9"
COMMENT     Sequence Label: IGF2BP1_2xRRM-4xKH_pGEX
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..723
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             733..846
                     /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(2503..2525)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2670..2688)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2756..2860
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2861..2929,2930..3721)
                     /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(3079..3098)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3892..4480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4381..4400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4634..4651
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4724..4801
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4802..5884
                     /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(4821..4840)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5933..5938,5939..5956,5957..5963)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5971..5987
                     /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     5995..6011
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5995..6011
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6007..6534
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6027..6044)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6027..6043)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6036..6058)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6636..6641,6642..6657,6658..6664)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggaagttct gttccagggg
      721 cccctgggat taatggacga aaaaaccacc ggttggcgtg gtggtcacgt tgttgaaggt
      781 ctggctggtg aactggaaca gctgcgtgct cgtctggaac accacccgca gggtcagcgt
      841 gaaccgggat ccatgaacaa gctttacatc ggcaacctca acgagagcgt gacccccgcg
      901 gacttggaga aagtgtttgc ggagcacaag atctcctaca gcggccagtt cttggtcaaa
      961 tccggctacg ccttcgtgga ctgcccggac gagcactggg cgatgaaggc catcgaaact
     1021 ttctccggga aagtagaatt acaaggaaaa cgcttagaga ttgaacattc ggtgcccaaa
     1081 aaacaaagga gccggaaaat tcaaatccga aatattccac cccagctccg atgggaagta
     1141 ctggacagcc tgctggctca gtatggtaca gtagagaact gtgagcaagt gaacaccgag
     1201 agtgagacgg cagtggtgaa tgtcacctat tccaaccggg agcagaccag gcaagccatc
     1261 atgaagctga atggccacca gttggagaac catgccctga aggtctccta catccccgat
     1321 gagcagatag cacagggacc tgagaatggg cgccgagggg gctttggctc tcggggtcag
     1381 ccccgccagg gctcacctgt ggcagcgggg gccccagcca agcagcagca agtggacatc
     1441 ccccttcggc tcctggtgcc cacccagtat gtgggtgcca ttattggcaa ggagggggcc
     1501 accatccgca acatcacaaa acagacccag tccaagatag acgtgcatag gaaggagaac
     1561 gcaggtgcag ctgaaaaagc catcagtgtg cactccaccc ctgagggctg ctcctccgct
     1621 tgtaagatga tcttggagat tatgcataaa gaggctaagg acaccaaaac ggctgacgag
     1681 gttcccctga agatcctggc ccataataac tttgtagggc gtctcattgg caaggaagga
     1741 cggaacctga agaaggtaga gcaagatacc gagacaaaaa tcaccatctc ctcgttgcaa
     1801 gaccttaccc tttacaaccc tgagaggacc atcactgtga agggggccat cgagaattgt
     1861 tgcagggccg agcaggaaat aatgaagaaa gttcgggagg cctatgagaa tgatgtggct
     1921 gccatgagcc tgcagtctca cctgatccct ggcctgaacc tggctgctgt aggtcttttc
     1981 ccagcttcat ccagcgcagt cccgccgcct cccagcagcg ttactggggc tgctccctat
     2041 agctccttta tgcaggctcc cgagcaggag atggtgcagg tgtttatccc cgcccaggca
     2101 gtgggcgcca tcatcggcaa gaaggggcag cacatcaaac agctctcccg gtttgccagc
     2161 gcctccatca agattgcacc acccgaaaca cctgactcca aagttcgtat ggttatcatc
     2221 actggaccgc cagaggccca attcaaggct cagggaagaa tctatggcaa actcaaggag
     2281 gagaacttct ttggtcccaa ggaggaagtg aagctggaga cccacatacg tgtgccagca
     2341 tcagcagctg gccgggtcat tggcaaaggt ggaaaaacgg tgaacgagtt gcagaatttg
     2401 acggcagctg aggtggtagt accaagagac cagacccctt gagcggccgc atcgtgactg
     2461 actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     2521 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2581 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     2641 cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2701 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2761 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2821 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2881 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2941 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3001 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3061 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     3121 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3181 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3241 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3301 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3361 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     3421 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3481 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3541 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3601 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3661 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3721 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3781 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3841 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3901 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3961 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4021 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4081 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4141 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4201 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4261 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4321 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4381 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4441 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4501 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4561 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4621 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     4681 tctccttacg catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg
     4741 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     4801 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     4861 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     4921 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     4981 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     5041 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     5101 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     5161 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     5221 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     5281 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     5341 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     5401 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     5461 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     5521 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     5581 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     5641 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     5701 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     5761 cgtctcactg gtgaaaagaa aaaccacgct agcgcccaat acgcaaaccg cctctccccg
     5821 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5881 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact
     5941 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     6001 acagctatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa
     6061 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6121 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6181 tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat
     6241 cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg
     6301 cccatctaca ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag
     6361 aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc
     6421 cagacgcgaa ttatttttga tggcgttgga attagcttat cgactgcacg gtgcaccaat
     6481 gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact
     6541 gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat
     6601 cataacggtt ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat
     6661 aatgtgtgga a
//
