LOCUS       sequence-220875-        5553 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human FUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5553)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-64: FUS-mEGFP
FEATURES             Location/Qualifiers
     source          1..5553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             997..1017
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1030..1074
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1114..1137
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1165..1167,1168..1881)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1207..1228)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1468..1487)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1815..1836
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2913..2929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2913..2929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2926..2948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2937..2953
                     /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(2961..2967,2968..2985,2986..2991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3006..3027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3144..3161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3315..3903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3395..3414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4074..4865,4866..4934))
                     /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     4697..4716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4935..5039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5107..5125
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5163..5185)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5285..5304
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5498..5520
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5512..5529
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5513..5529
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgctctttga tatattggta ctgaggtatg tgcgtgtttt ccaaagaagt aaatgtcaag
       61 gccacactgt tggggtcaga tttagccaaa agcttaccta ggtaaggttg atgtaatggg
      121 aaaggtaatg gattgggttc agtaatactg atttttgttc ctgactctga gaagcaagcc
      181 gttttgtctt tctgaagctt cagtttcctc actgtatctc taaagtcacc gtagtttctt
      241 cctagttcta ggtcttgcct attccccatc gctccagact gattgtcttc ctttctcctt
      301 agcacctgtg agaatatgaa cttctcttgg aggaatgaat gcaaccagtg taaggcccct
      361 aaaccagatg gcccaggagg gggaccaggt ggctctcaca tgggtaagaa aggcagacct
      421 ggtgctaggg agctgggacc aaagaatcct taatttttca gcggggaggc tcggggaaca
      481 taggggaatg ggaatatgat agatcttgtt tcttttgtcc tagggggtaa ctacggggat
      541 gatcgtcgtg gtggcagagg aggctatgat cgaggcggct accggggccg cggcggggac
      601 cgtggaggct tccgaggggg ccggggtggt ggggacagag gtggctttgg ccctggcaag
      661 atggattcca ggtaagactt taaatcagaa taaaaaagta gagcagttga acagaggcca
      721 taggataaca gggttttgtt gagaaagtgg tttcattttg agggctaggt ggaaagacct
      781 gaggttgtaa ccagtagtgg agagggaagg aaaattaact cagggggagt gaatctgtag
      841 acccacttga gataagatac tcgctgggtt aggtaggagg ggcagatagg atatctaggc
      901 ttggagaggc tggtaactca aatataatgg atacttaatt tttttttttt tttttgcagg
      961 ggtgagcaca gacaggatcg cagggagcgc ccgtatgaga atctttattt tcagggcgcc
     1021 gccaaattca aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga cagcggaggt
     1081 ggaggttcga gcggtccctc gggttcgtcg agcctggaag ttctgttcca ggggcccctg
     1141 tcgtcgagcg gtccctcggg ttcggtgagc aagggcgagg agctgttcac cggggtggtg
     1201 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1261 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1321 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1381 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1441 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1501 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1561 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1621 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1681 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1741 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     1801 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1861 atggacgagc tgtacaagta attagactgg ctccccaggt tctggaacag ctttttgtcc
     1921 tgtacccagt gttaccctcg ttattttgta accttccaat tcctgatcac ccaagggttt
     1981 ttttgtgtcg gactatgtaa ttgtaactat acctctggtt cccattaaaa gtgaccattt
     2041 tagttaaatt ttgttcctct tccccctttt cactttcctg gaagatcgat gtcccgatca
     2101 ggaaggtaga gagttttcct gttcagatta ccctgcccag caggaactgg aatacagtgt
     2161 tcggggagaa ggccaaatga tatccttgag agcagagatt aaacttttct gtcatgggga
     2221 aagttggtgt ataaatgaga aatgaagaac atgggatgtc atgagtgttg gcctaaattt
     2281 gcccagctat ggggaatttt tcctttacca catttatttg catactggtc ttagtttatt
     2341 tgcagcagtt tatccctttt taagaactct ttgatctttt ggccctttta atggtgaggc
     2401 tcaaacaaac tacatttaaa tggggcagta ttcagatttg accatggtgg agagcgctta
     2461 gccactctgg gtctttcaca ggaaggagag taactgagtg ctgcaggagt ttgtggagtg
     2521 gagtcaggat ctaggaggtg agtgactccc ttcctagctg ccctggtgaa cagcgcttgg
     2581 gtagatacct gctataagga gactggtctg gctgggttac tttcacatcc tgcctgtact
     2641 cagagggctt caggtcattg acattatgag attttaggct tgatcccttt ttgattggag
     2701 ggtggaaggc cctcctaagg gaatgataag tgataagagg gggaaggggt tgcagccaat
     2761 gagttaaaac cttagagcag tgctcctcag cctcttacca tgtggttgta aacttgcacg
     2821 tacctgccaa ccagttattt agcatgcttt ttattttagt tacacagagc gtaacattat
     2881 cccaatggcg cgccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3721 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5041 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5101 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5161 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5221 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5281 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5341 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     5401 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5461 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5521 acggccagtg aattgacgcg tattgggatt gcg
//
