LOCUS       sequence-274844-        6183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express HRP-1 with a C-terminal maltose binding protein tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6183)
  AUTHORS   Ryan VH, Perdikari TM, Naik MT, Saueressig CF, Lins J, Dignon GL, 
            Mittal J, Hart AC, Fawzi NL
  TITLE     Tyrosine phosphorylation regulates hnRNPA2 granule protein 
            partitioning and reduces neurodegeneration.
  JOURNAL   EMBO J. 2021 Feb 1;40(3):e105001. doi: 10.15252/embj.2020105001. 
            Epub 2020 Dec 22.
  PUBMED    33349959
REFERENCE   2  (bases 1 to 6183)
  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: "EMBO J."; 
            date: "2021-02-1"; volume: "40(3):e105001. doi"; pages: " 
            10.15252/embj.2020105001. Epub 2020 Dec 22"
COMMENT     Sequence Label: pJ4M_HRP-1_FL
FEATURES             Location/Qualifiers
     source          1..6183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             267..287
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             297..1394
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1251..1260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1368..1391
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1413..1437
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1446..1463
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1471..1518
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1741..1760)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1827..1918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1919..2728
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2811..3893
                     /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(2830..2849)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3979..4567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4468..4487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4721..4738
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4836..4853
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4837..4853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5317..5336
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5317..5335
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5336..5360
                     /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)."
ORIGIN
        1 ggggtcagca aggtggtggt ggccagggtg gttggggtgg tccgcagcag caacagggtg
       61 gtggtgggtg gggtcaacaa ggcggtggcg gtcaaggtgg atggggtggc ccacagcaac
      121 aacagcaagg cggttgggga ggtccgcaac aaggtggcgg cggtggaggt tggggcggtc
      181 agggtcagca gcagggtggc tggggtggtc aatctggtgc gcaacagtgg gctcatgcgc
      241 agggcggcaa tcgtaactat ctcgaggaga acctgtactt tcagggtagc ggtaccaaaa
      301 tcgaagaagg taaactggta atctggatta acggcgataa aggctataac ggtctcgctg
      361 aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata
      421 aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac attatcttct
      481 gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg
      541 acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca
      601 agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc
      661 tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga
      721 aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg
      781 ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg
      841 gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca
      901 aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa
      961 cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt
     1021 atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc
     1081 tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa
     1141 actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg
     1201 tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccaccatgg
     1261 aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg
     1321 ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga
     1381 aagacgcgca gactaattcg ggatctgata tcacaagttt gtacaaaaaa gctgaaggcg
     1441 gtacccatca ccatcaccat cattgacccc ctagcataac cccttggggc ctctaaacgg
     1501 gtcttgaggg gttttttgcc cctgagacgc gtcaatcgag ttcgtaccta agggcgacac
     1561 cccctaatta gcccgggcga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     1621 gcctggcagt tccctactct cgcatgggga gtccccacac taccatcggc gctacggcgt
     1681 ttcacttctg agttcggcat ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa
     1741 caaggggtgt tatgagccat attcaggtat aaatgggctc gcgataatgt tcagaattgg
     1801 ttaattggtt gtaacactga cccctatttg tttatttttc taaatacatt caaatatgta
     1861 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatat
     1921 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     1981 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2041 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2101 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc cacttccgac
     2161 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2221 aaaaacagcg ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2281 gctggcagtg ttcctgcgcc ggttgcactc gattcctgtt tgtaattgtc cttttaacag
     2341 cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2401 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2461 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2521 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2581 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     2641 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcaatt
     2701 tcatttgatg ctcgatgagt ttttctaagc ggcgcgccat cgaatggcgc aaaacctttc
     2761 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat atgaaaccag
     2821 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     2881 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     2941 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     3001 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     3061 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     3121 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     3181 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     3241 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     3301 aggacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     3361 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     3421 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     3481 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     3541 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     3601 ttggtgcgga tatctcggta gtgggatacg acgataccga agatagctca tgttatatcc
     3661 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     3721 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgcca gtctcactgg
     3781 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3841 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgactcatga
     3901 ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga
     3961 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4021 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4081 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4141 gtagttagcc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4201 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4261 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4321 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4381 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4441 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4501 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4561 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4621 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4681 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4741 agcggaaggc gagagtaggg aactgccagg catcaaacta agcagaaggc ccctgacgga
     4801 tggccttttt gcgtttctac aaactctttc tgtgttgtaa aacgacggcc agtcttaagc
     4861 tcgggccccc tgggcggttc tgataacgag taatcgttaa tccgcaaata acgtaaaaac
     4921 ccgcttcggc gggttttttt atggggggag tttagggaaa gagcatttgt cagaatattt
     4981 aagggcgcct gtcactttgc ttgatatatg agaattattt aaccttataa atgagaaaaa
     5041 agcaacgcac tttaaataag atacgttgct ttttcgattg atgaacacct ataattaaac
     5101 tattcatcta ttatttatga ttttttgtat atacaatatt tctagtttgt taaagagaat
     5161 taagaaaata aatctcgaaa ataataaagg gaaaatcagt ttttgatatc aaaattatac
     5221 atgtcaacga taatacaaaa tataatacaa actataagat gttatcagta tttattatgc
     5281 atttagaata aattttgtgt cgcccttccg cgaaattaat acgactcact ataggggaat
     5341 tgtgagcgga taacaattcc cctctagaaa taattttgtt taacttttga aggagatata
     5401 cccatatgac cgacgtcgag attaaagccg agaacggctc gggtgatgcc tctctggagc
     5461 cggagaatct gcgcaagatc ttcgttggcg gcctgacgag caataccacc gatgacctga
     5521 tgcgtgagtt ctacagccaa tttggcgaga tcaccgacat tattgttatg cgcgatccga
     5581 ctacgaagcg cagccgtggc tttggctttg tgacgtttag cggcaagact gaagttgacg
     5641 ctgcgatgaa gcaacgcccg catatcattg acggcaagac cgtggacccg aagcgtgcag
     5701 ttccgcgtga tgacaaaaac cgcagcgaaa gcaacgttag caccaaacgc ctgtacgtga
     5761 gcggtgtgcg tgaagatcac accgaagata tgctgaccga gtacttcacc aaatatggta
     5821 ccgtcacgaa aagcgagatt atcttggata aagcgacgca aaagccgcgt ggtttcggct
     5881 ttgtcacctt cgacgatcac gattccgttg accagtgtgt gctgcagaaa agccacatgg
     5941 tgaatggcca tcgctgcgac gtccgtaagg gtttgagcaa agacgaaatg tccaaggcac
     6001 aaatgaaccg tgacagagaa acgcgcggtg gtcgtagccg tgatggtcag cgtggcggct
     6061 acaacggtgg tggcggtggt ggtggaggat ggggcggtcc agcacagcgt ggtggccctg
     6121 gtgcgtacgg cggtcctggt ggtgggggtc agggtggcta tggcggcgat tatggtggcg
     6181 gct
//
