LOCUS       sequence-455751-        7282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human progranulin (PGRN) with an N-terminal twin-Strep-V5 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7282)
  AUTHORS   Root J, Mendsaikhan A, Taylor G, Merino P, Nandy S, Wang M, Araujo 
            LT, Ryu D, Holler C, Thompson BM, Astarita G, Blain JF, Kukar T
  TITLE     Granulins rescue inflammation, lysosome dysfunction, lipofuscin, and
            neuropathology in a mouse model of progranulin deficiency.
  JOURNAL   Cell Rep. 2024 Nov 19;43(12):114985. doi: 
            10.1016/j.celrep.2024.114985.
  PUBMED    39565694
REFERENCE   2  (bases 1 to 7282)
  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: "Cell Rep.";
            date: "2024-11-19"; volume: "43(12):114985. doi"; pages: " 
            10.1016/j.celrep.2024.114985"
COMMENT     Sequence Label: pAAV hPGRN
FEATURES             Location/Qualifiers
     source          1..7282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     promoter        439..716
                     /label=chicken beta-actin promoter
     primer_bind     995..1017
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1079..1098
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1194..1199,1200..1223,1224..1259,1260..1283)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             1293..1334
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1335..1355
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    3105..3693
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3158..3178)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3576..3587)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3720..3737)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3726..3950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4160..4489)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4174..4309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4474..4494
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4735..5323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4815..4834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5494..6285,6286..6354))
                     /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     6117..6136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6355..6459)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6527..6545
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6583..6605)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6705..6724
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6938..6957
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7247..7265
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7247..7264
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      241 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      301 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      361 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      421 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      481 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      541 gggcgggggg gggggggggg sgcgcgccgg gggggggggg ggggggsggg gggsggggsg
      601 gggsgrggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      661 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      721 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      781 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      841 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      901 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
      961 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1021 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1081 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatca aagcttgcca
     1141 ccatgtggac cctggtgtcc tgggtggccc tgacagccgg actggtggcc ggatctgcct
     1201 ggtcccaccc ccagtttgag aagggcggag gctctggcgg cggaagcgga ggatctgctt
     1261 ggagccaccc tcagttcgaa aagggggcca gcggcaagcc catccccaat cctctgctgg
     1321 gcctggacag caccgagaac ctgtattttc aaggcacccg gtgtcccgac ggccagtttt
     1381 gccctgtggc ctgctgcctg gaccctggcg gagccagcta cagctgctgc agacccctgc
     1441 tggacaagtg gcccaccacc ctgagcagac acctgggcgg accttgccag gtggacgccc
     1501 actgttctgc cggccacagc tgcatcttca ccgtgtccgg cacctccagc tgctgcccat
     1561 ttcctgaggc cgtggcctgt ggcgacggac accactgctg ccctagaggc ttccactgca
     1621 gcgccgacgg cagaagctgc ttccagcgga gcggcaacaa cagcgtgggc gccatccagt
     1681 gccccgactc ccagttcgag tgccccgatt tcagcacctg ttgcgtgatg gtggacggca
     1741 gctggggctg ctgtcccatg cctcaggcca gctgttgcga ggaccgggtg cactgttgcc
     1801 ctcacggcgc cttttgcgac ctggtgcaca cccggtgcat cacccctacc ggcacccacc
     1861 ctctggccaa gaaactgccc gcccagcgga ccaacagagc cgtggccctg tctagcagcg
     1921 tgatgtgccc cgatgccaga agccgctgcc ctgacggcag cacatgctgc gagctgccca
     1981 gcgggaagta cggctgttgc ccaatgccca acgccacctg ttgcagcgac catctgcact
     2041 gctgtccaca agacaccgtg tgcgacctga tccagagcaa gtgcctgagc aaagagaacg
     2101 ccaccaccga cctgctgacc aagctgcctg cccacaccgt gggcgacgtg aagtgcgaca
     2161 tggaagtgtc ctgccccgat ggctacacct gttgccggct gcagagcggc gcctggggat
     2221 gctgcccttt cacccaggcc gtgtgttgtg aggaccacat ccattgctgc cctgccggct
     2281 tcacctgtga cacccagaag ggaacctgcg agcagggccc acaccaggtg ccctggatgg
     2341 aaaaggcccc tgcccacctg agcctgcccg atcctcaggc cctgaagagg gacgtgccct
     2401 gcgacaacgt gtccagctgc cccagcagcg atacctgctg ccagctgaca tccggcgagt
     2461 ggggctgttg tcctatcccc gaagccgtgt gctgctccga tcaccagcac tgttgtcctc
     2521 aaggctacac atgcgtggcc gagggccagt gtcagcgggg atctgagatc gtggccggcc
     2581 tggaaaagat gcccgccaga agggccagcc tgagccaccc cagagacatc ggctgcgacc
     2641 agcacacctc ctgccctgtg ggccagactt gctgtcctag cctgggcggc tcctgggcct
     2701 gctgtcagct gcctcacgct gtgtgctgtg aggataggca gcattgctgt cccgccggat
     2761 acacatgcaa cgtgaaggcc cggtcctgcg agaaagaagt ggtgtccgcc cagcccgcca
     2821 ccttcctggc cagatctcca cacgtgggag tgaaggacgt ggaatgcggc gagggccact
     2881 tctgccacga caaccagaca tgctgccggg acaaccgcca gggctgggcc tgttgcccat
     2941 acagacaggg cgtgtgctgt gctgatcggc gccactgttg cccagccggc tttagatgcg
     3001 ccgccagggg caccaagtgt ctgcggagag aagcccccag atgggacgcc cccctgagag
     3061 atcccgccct gagacagctg ctgtgactcg agcagcttat cgataatcaa cctctggatt
     3121 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3181 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3241 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3301 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3361 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3421 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3481 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     3541 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3601 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3661 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgactc gctgatcagc
     3721 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3781 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3841 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3901 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3961 ggaaagaacc agctggggct cgactagagc atggctacgt agataagtag catggcgggt
     4021 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4081 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc
     4141 gagcgagcgc gcagagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     4201 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     4261 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     4321 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     4381 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     4441 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     4501 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4561 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4621 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6421 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6481 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6541 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6601 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6661 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6721 tgtactgaga gtgcaccatt cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     6781 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     6841 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     6901 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6961 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     7021 ggctagcgat gaccctgctg attggttcgc tgaccatttc cgggtgcggg acggcgttac
     7081 cagaaactca gaaggttcgt ccaaccaaac cgactctgac ggcagtttac gagagagatg
     7141 atagggtctg cttcagtaag ccagatgcta cacaattagg cttgtacata ttgtcgttag
     7201 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     7261 atagaataca cggaattaat tc
//
