LOCUS       sequence-274711-        7168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mScarlet tagged hrp-1HsLCWT in mec-4 neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7168)
  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 7168)
  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: pHA848 - mec-4p::hrp-1HsLCWTmScarlet
FEATURES             Location/Qualifiers
     source          1..7168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..91
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     83..100
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     84..100
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     110..129
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        110..128
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2010..2705
                     /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(2143..2161)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2411..2430
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4418..4434)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4419..4435)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4464..4484)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4464..4482)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4503..4519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4503..4519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4516..4538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4527..4543
                     /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(4551..4557,4558..4575,4576..4581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4596..4617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4734..4751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4905..5493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4985..5004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5664..6455,6456..6524))
                     /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     6287..6306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6525..6629)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6656..7111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6743..6762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6953..6974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
       61 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
      121 actatagggc gaattggagc tccaccgcgg tggcggccgc taagcttcaa tacaagctca
      181 aatacaatac tatttttttt ccgtctgaac ataaaattag tttctaaata aaaaatgtga
      241 aaataaatat ttggttttaa tcgttaaaat cttccaaatt actaataaag ttcagtcaac
      301 tatatgcttc gaaattcact gaatctatta ttgttattaa tttttagacc cttggagagc
      361 tcccaactct ttcaatccag aagactgatg gatgccaggt gtacttgtca aaggtggctc
      421 aaggatgcga gattgtcact tccaaatcca gtgagatgaa catctccgtg caaaccaatg
      481 atgatgggga ttatgtaagt ttttcggatt gtagaaccca atctaaattt ccttttcagt
      541 cggagttccc ggttccggag cagttcaaga caacttttgt caatggcaag ctagtcactg
      601 tcgtctcgga catttgctaa atcttctcac gtcataacct tatttttcaa cttcatgaac
      661 tatcccctcg aactttttta tttgtccctg tctacaattc tttataagca attgagacac
      721 actttgcttt attaaataat accgattttg ttttgtatat gccttgactg gtgctcaact
      781 atttttattg aaaaaattac tttttactgt ctcgtcgaat aaaagtgatc ccttccaaaa
      841 aatagttatt catgtttgaa ggtttgtgat cgtagaacaa ttgcaccgaa tgtagtgtaa
      901 aaaaaattcc agaatgttca aatacatttc caccctcacc agaacatctc aaacaccagg
      961 cacatttcaa cacactttca tggatcttat cttgcctcaa aaccatcagc tttcttctca
     1021 ttctacaagt atgaaacaat ctactattct atacatttat aatgtaatgg aaccatgtga
     1081 agacatgagc aagatcattg tcgtcagcga ctacccctcc ttcatttttc ttagagtatc
     1141 agtttgctaa tgctattttt tttatcgcta tcaagttata gagggatccc cgggattggc
     1201 caaaggaccc aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg
     1261 acccttgctt ggagggtacc ggtagaaaaa atgacggacg tggaaatcaa ggctgaggtt
     1321 acgtttgttc acttgctttt agggtgtagg ccacctgatt tgggtcccga tcatttaatt
     1381 atcgatttta ttttcagaat ggaagcggag atgcctcgct cgagccagag aaccttcgaa
     1441 agattttcgt tggaggactc acgtcaaaca ccaccgatga cctgatgcga gagttctact
     1501 cacaattcgg agaaatcact gatatcatcg tcatgagaga tccaactacc aagagatccc
     1561 gcggattcgg atttgtcacc ttctctggca aaactgaagt cgatgctgcc atgaaacaac
     1621 gcccgcacat catcgacgga aagaccgtgg acccgaagcg tgccgtgcca cgtgatgata
     1681 agaaccgctc cgagtcgaat gtctccacca aacgcctgta cgtcagcgga gttcgtgagg
     1741 atcacaccga ggacatgctc accgagtatt tcacaaagta tggaaccgtc accaaatccg
     1801 agattattct cgacaaagcc acccaaaagc caagaggctt cggattcgtc accttcgatg
     1861 atcatgactc tgtggatcaa tgcgttcttc aaaaatccca tatggtcaac ggacacagat
     1921 gtgacgtgcg caaaggactc tccaaggacg agatgagcaa ggctcagatg aaccgcgatc
     1981 gtgaaacccg tggcggaaga tctcgcgata tggtgagcaa gggcgaggca gtgatcaagg
     2041 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     2101 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2161 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2221 gggccttcac caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
     2281 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     2341 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     2401 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc accgagcggt
     2461 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     2521 gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     2581 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     2641 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     2701 acaagggaca gcgtggcggt tacaatggcg gtggtggtgg cggcggcggc tggggaggac
     2761 cagctcaacg cggcggtcca ggagcatatg gtggaccagg aggtggcggc caaggaggat
     2821 atggcggagg tagttggatt ctggcttgtg aattctcgat tctaatatta ttaactttca
     2881 gattacggag gaggacgcgg tggttatggt ggtggaggac caggatatgg aaatcagggt
     2941 ggaggatatg gaggaggata tgataattat ggaggaggaa attacggatc tggaaattac
     3001 aatgatttcg gaaactacaa ccagcagcca tctaattatg gaccaatgaa gtctggaaat
     3061 ttcggaggat ctcgcaatat gggtggacca tatggaggag gaaattatgg tccaggagga
     3121 tctggaggat ctggaggata tggaggacgt tctcgttact aaattaattc cttaagcccc
     3181 tctaagtgtc caacgtgtct gagcttcccg tgctctctcc tctgtatcga tcttctcaac
     3241 catttttgct ctgctatctc atcaaaatct tctgtacttt ttcttgttgc ctttctccag
     3301 aaagtttcgt ttattattca attttttacc ctgtcttttt tatataattt atttctaaat
     3361 tcgcacagtt gacatccatt caactagaga atgaatgcag agtttgacac caaattggct
     3421 taattttcaa gctgttttta accaaaataa taaaaaagtc tcaactttac aaaaatgttt
     3481 atctggatct ggacgacagc gagctgagaa attgatatat tttcagagta aaaactaaaa
     3541 atgtctggaa tttttggagt cattgtctca gggaggacgc cgatcgaagt ggtgccggtc
     3601 tccgatacgg agttcacctg cgaaattgta aacgctgacg cgataaacca cgtggtggtc
     3661 ttccttaccg gagctcaacc gtttccggac ggaattggtg gctctggtga tcattttcag
     3721 aattttccgc ccaaaaaaac ccaatttatt tcagtttaca tcagatggcc cacccaagac
     3781 ggcggaaatt ggcattattt gggatttatt tgcaacgaga agcccagcgc gatttataaa
     3841 gttgctcagg tgtgcatttt tgatggaaaa atgaaagaaa accaccgaaa tttcgatgtt
     3901 tctctgtcga ttttttttat ttaaaaactt agaaatactc tcaatttcgc atttttcacc
     3961 aaaaaaaaga atctcaactg gtttttcaca tagcttaatt tttgaagaaa aatctcaaca
     4021 aaacccagtt ttaagtttca aattctcaaa aattgatctt ttttttccct taaaaatcca
     4081 gtttttagct ccaaattgtc ataattttcc agaaaaattt agttttcgag ctaacaacat
     4141 taaaatttca cagaaaatct gaatttcagc cttagagatc caaatttttc ggctgaaaat
     4201 cacaattttt ccatttttca gctccacaaa tccgacgctt ctcactcgtt ttcccaattt
     4261 gacaatcaaa tgcaactata ttccagtgga tcagctcaaa ttggaattaa tgctgaaagt
     4321 ttatctgata tcactggacg gcaggcagct gatgggacac aggctagcca gcaggtgaag
     4381 gatcccccgg gctgcaggaa ttcgatatca agcttatcga taccgtcgac ctcgaggggg
     4441 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     4501 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5341 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5941 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6541 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6601 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     6661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6961 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     7021 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7081 aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt
     7141 gggaagggcg atcggtgcgg gcctcttc
//
