LOCUS       sequence-274709-        7165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mScarlet tagged hrp-1 in mec-4 neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7165)
  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 7165)
  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: pHA847 - mec-4p::hrp-1mScarlet
FEATURES             Location/Qualifiers
     source          1..7165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..88
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     80..97
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     81..97
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     107..126
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        107..125
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2007..2702
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2140..2158)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2408..2427
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4412..4428)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4413..4429)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4458..4478)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4458..4476)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4497..4513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4497..4513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4510..4532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4521..4537
                     /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)."
     promoter        complement(join(4545..4551,4552..4569,4570..4575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4590..4611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4728..4745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4899..5487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4979..4998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5658..6449,6450..6518))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6281..6300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6519..6623)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6650..7105
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6737..6756)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6947..6968
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
       61 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
      121 atagggcgaa ttggagctcc accgcggtgg cggccgctaa gcttcaatac aagctcaaat
      181 acaatactat tttttttccg tctgaacata aaattagttt ctaaataaaa aatgtgaaaa
      241 taaatatttg gttttaatcg ttaaaatctt ccaaattact aataaagttc agtcaactat
      301 atgcttcgaa attcactgaa tctattattg ttattaattt ttagaccctt ggagagctcc
      361 caactctttc aatccagaag actgatggat gccaggtgta cttgtcaaag gtggctcaag
      421 gatgcgagat tgtcacttcc aaatccagtg agatgaacat ctccgtgcaa accaatgatg
      481 atggggatta tgtaagtttt tcggattgta gaacccaatc taaatttcct tttcagtcgg
      541 agttcccggt tccggagcag ttcaagacaa cttttgtcaa tggcaagcta gtcactgtcg
      601 tctcggacat ttgctaaatc ttctcacgtc ataaccttat ttttcaactt catgaactat
      661 cccctcgaac ttttttattt gtccctgtct acaattcttt ataagcaatt gagacacact
      721 ttgctttatt aaataatacc gattttgttt tgtatatgcc ttgactggtg ctcaactatt
      781 tttattgaaa aaattacttt ttactgtctc gtcgaataaa agtgatccct tccaaaaaat
      841 agttattcat gtttgaaggt ttgtgatcgt agaacaattg caccgaatgt agtgtaaaaa
      901 aaattccaga atgttcaaat acatttccac cctcaccaga acatctcaaa caccaggcac
      961 atttcaacac actttcatgg atcttatctt gcctcaaaac catcagcttt cttctcattc
     1021 tacaagtatg aaacaatcta ctattctata catttataat gtaatggaac catgtgaaga
     1081 catgagcaag atcattgtcg tcagcgacta cccctccttc atttttctta gagtatcagt
     1141 ttgctaatgc tatttttttt atcgctatca agttatagag ggatccccgg gattggccaa
     1201 aggacccaaa ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc
     1261 cttgcttgga gggtaccggt agaaaaaatg acggacgtgg aaatcaaggc tgaggttacg
     1321 tttgttcact tgcttttagg gtgtaggcca cctgatttgg gtcccgatca tttaattatc
     1381 gattttattt tcagaatgga agcggagatg cctcgctcga gccagagaac cttcgaaaga
     1441 ttttcgttgg aggactcacg tcaaacacca ccgatgacct gatgcgagag ttctactcac
     1501 aattcggaga aatcactgat atcatcgtca tgagagatcc aactaccaag agatcccgcg
     1561 gattcggatt tgtcaccttc tctggcaaaa ctgaagtcga tgctgccatg aaacaacgcc
     1621 cgcacatcat cgacggaaag accgtggacc cgaagcgtgc cgtgccacgt gatgataaga
     1681 accgctccga gtcgaatgtc tccaccaaac gcctgtacgt cagcggagtt cgtgaggatc
     1741 acaccgagga catgctcacc gagtatttca caaagtatgg aaccgtcacc aaatccgaga
     1801 ttattctcga caaagccacc caaaagccaa gaggcttcgg attcgtcacc ttcgatgatc
     1861 atgactctgt ggatcaatgc gttcttcaaa aatcccatat ggtcaacgga cacagatgtg
     1921 acgtgcgcaa aggactctcc aaggacgaga tgagcaaggc tcagatgaac cgcgatcgtg
     1981 aaacccgtgg cggaagatct cgcgatatgg tgagcaaggg cgaggcagtg atcaaggagt
     2041 tcatgcggtt caaggtgcac atggagggct ccatgaacgg ccacgagttc gagatcgagg
     2101 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     2161 gtggccccct gcccttctcc tgggacatcc tgtcccctca gttcatgtac ggctccaggg
     2221 ccttcaccaa gcaccccgcc gacatccccg actactataa gcagtccttc cccgagggct
     2281 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg acccaggaca
     2341 cctccctgga ggacggcacc ctgatctaca aggtgaagct ccgcggcacc aacttccctc
     2401 ctgacggccc cgtaatgcag aagaagacaa tgggctggga agcgtccacc gagcggttgt
     2461 accccgagga cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg aaggacggcg
     2521 gccgctacct ggcggacttc aagaccacct acaaggccaa gaagcccgtg cagatgcccg
     2581 gcgcctacaa cgtcgaccgc aagttggaca tcacctccca caacgaggac tacaccgtgg
     2641 tggaacagta cgaacgctcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2701 agggacagcg tggcggttac aatggcggtg gtggtggcgg cggcggctgg ggaggaccag
     2761 ctcaacgcgg cggtccagga gcatatggtg gaccaggagg tggcggccaa ggaggatatg
     2821 gcggaggtag ttggattctg gcttgtgaat tctcgattct aatattatta actttcagat
     2881 tacggcggtg gctggggaca gcaaggaggc ggcggtcaag gcggatgggg aggcccacaa
     2941 cagcagcaag gaggtggtgg ctggggacag caaggcggcg gcggtcaagg cggatgggga
     3001 ggaccacagc aacagcagca aggaggctgg ggaggtcctc aacaaggagg cggcggcggt
     3061 ggctggggag gccaaggcca gcagcaaggc ggctggggag gtcaatccgg tgctcagcag
     3121 tgggctcatg ctcaaggagg aaacagaaac tattaaatta attccttaag cccctctaag
     3181 tgtccaacgt gtctgagctt cccgtgctct ctcctctgta tcgatcttct caaccatttt
     3241 tgctctgcta tctcatcaaa atcttctgta ctttttcttg ttgcctttct ccagaaagtt
     3301 tcgtttatta ttcaattttt taccctgtct tttttatata atttatttct aaattcgcac
     3361 agttgacatc cattcaacta gagaatgaat gcagagtttg acaccaaatt ggcttaattt
     3421 tcaagctgtt tttaaccaaa ataataaaaa agtctcaact ttacaaaaat gtttatctgg
     3481 atctggacga cagcgagctg agaaattgat atattttcag agtaaaaact aaaaatgtct
     3541 ggaatttttg gagtcattgt ctcagggagg acgccgatcg aagtggtgcc ggtctccgat
     3601 acggagttca cctgcgaaat tgtaaacgct gacgcgataa accacgtggt ggtcttcctt
     3661 accggagctc aaccgtttcc ggacggaatt ggtggctctg gtgatcattt tcagaatttt
     3721 ccgcccaaaa aaacccaatt tatttcagtt tacatcagat ggcccaccca agacggcgga
     3781 aattggcatt atttgggatt tatttgcaac gagaagccca gcgcgattta taaagttgct
     3841 caggtgtgca tttttgatgg aaaaatgaaa gaaaaccacc gaaatttcga tgtttctctg
     3901 tcgatttttt ttatttaaaa acttagaaat actctcaatt tcgcattttt caccaaaaaa
     3961 aagaatctca actggttttt cacatagctt aatttttgaa gaaaaatctc aacaaaaccc
     4021 agttttaagt ttcaaattct caaaaattga tctttttttt cccttaaaaa tccagttttt
     4081 agctccaaat tgtcataatt ttccagaaaa atttagtttt cgagctaaca acattaaaat
     4141 ttcacagaaa atctgaattt cagccttaga gatccaaatt tttcggctga aaatcacaat
     4201 ttttccattt ttcagctcca caaatccgac gcttctcact cgttttccca atttgacaat
     4261 caaatgcaac tatattccag tggatcagct caaattggaa ttaatgctga aagtttatct
     4321 gatatcactg gacggcaggc agctgatggg acacaggcta gccagcaggt gaaggatccc
     4381 ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg
     4441 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     4501 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4561 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4621 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4681 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4741 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4801 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5521 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5581 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5641 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5701 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5761 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5821 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5881 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5941 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6001 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6061 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6121 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6181 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6241 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6301 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6361 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6421 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6481 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6541 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6601 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     6661 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6721 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6781 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6841 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6901 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6961 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     7021 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     7081 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     7141 ggcgatcggt gcgggcctct tcgct
//
