LOCUS       sequence-469050-        8584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing HA-tagged human progranulin that is 
            fused to the transmembrane domain and cytosolic tail of LAMP-1. 
            Enables lysosomal delivery of progranulin without secretion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8584)
  AUTHORS   Davis SE, Roth JR, Aljabi Q, Hakim AR, Savell KE, Day JJ, Arrant AE
  TITLE     Delivering progranulin to neuronal lysosomes protects against 
            excitotoxicity.
  JOURNAL   J Biol Chem. 2021 Sep;297(3):100993. doi: 10.1016/j.jbc.2021.100993.
            Epub 2021 Jul 21.
  PUBMED    34298019
REFERENCE   2  (bases 1 to 8584)
  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: "J Biol 
            Chem."; date: "2021-09"; volume: "297(3):100993. doi"; pages: " 
            10.1016/j.jbc.2021.100993. Epub 2021 Jul 21"
COMMENT     Sequence Label: PGK-HA-PGRN-LAMP1TM
FEATURES             Location/Qualifiers
     source          1..8584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1972..2482
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2401..2419
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2551..2577
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    4435..5023
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4488..4508)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4906..4917)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5110..5343
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5415..5536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5452..5471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5506..5525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5576..5711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5731..5750)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5732..5750)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5760..5777)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5760..5776)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5769..5791)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5918..6373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6005..6024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6215..6236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6399..6503
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6504..6572,6573..7364)
                     /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     complement(6722..6741)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7535..8123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8024..8043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8277..8294
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8411..8432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8447..8452,8453..8470,8471..8477)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8485..8501
                     /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)."
     primer_bind     8490..8512
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8509..8525
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8509..8525
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8544..8564
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8546..8564
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttatcg atcacgagac tagcctcgag aagcttgata tcgaattcca cggggttggg
     1981 gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
     2041 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
     2101 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
     2161 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
     2221 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
     2281 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
     2341 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
     2401 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
     2461 aatcaccgac ctctctcccc agggggatcc gccaccatgt ggaccctggt gagctgggtg
     2521 gccttaacag cagggctggt ggctggaggt tacccatacg atgttccaga ttacgctggt
     2581 acgcggtgcc cagatggtca gttctgccct gtggcctgct gcctggaccc cggaggagcc
     2641 agctacagct gctgccgtcc ccttctggac aaatggccca caacactgag caggcatctg
     2701 ggtggcccct gccaggttga tgcccactgc tctgccggcc actcctgcat ctttaccgtc
     2761 tcagggactt ccagttgctg ccccttccca gaggccgtgg catgcgggga tggccatcac
     2821 tgctgcccac ggggcttcca ctgcagtgca gacgggcgat cctgcttcca aagatcaggt
     2881 aacaactccg tgggtgccat ccagtgccct gatagtcagt tcgaatgccc ggacttctcc
     2941 acgtgctgtg ttatggtcga tggctcctgg gggtgctgcc ccatgcccca ggcttcctgc
     3001 tgtgaagaca gggtgcactg ctgtccgcac ggtgccttct gcgacctggt tcacacccgc
     3061 tgcatcacac ccacgggcac ccaccccctg gcaaagaagc tccctgccca gaggactaac
     3121 agggcagtgg ccttgtccag ctcggtcatg tgtccggacg cacggtcccg gtgccctgat
     3181 ggttctacct gctgtgagct gcccagtggg aagtatggct gctgcccaat gcccaacgcc
     3241 acctgctgct ccgatcacct gcactgctgc ccccaagaca ctgtgtgtga cctgatccag
     3301 agtaagtgcc tctccaagga gaacgctacc acggacctcc tcactaagct gcctgcgcac
     3361 acagtggggg atgtgaaatg tgacatggag gtgagctgcc cagatggcta tacctgctgc
     3421 cgtctacagt cgggggcctg gggctgctgc ccttttaccc aggctgtgtg ctgtgaggac
     3481 cacatacact gctgtcccgc ggggtttacg tgtgacacgc agaagggtac ctgtgaacag
     3541 gggccccacc aggtgccctg gatggagaag gccccagctc acctcagcct gccagaccca
     3601 caagccttga agagagatgt cccctgtgat aatgtcagca gctgtccctc ctccgatacc
     3661 tgctgccaac tcacgtctgg ggagtggggc tgctgtccaa tcccagaggc tgtctgctgc
     3721 tcggaccacc agcactgctg cccccagggc tacacgtgtg tagctgaggg gcagtgtcag
     3781 cgaggaagcg agatcgtggc tggactggag aagatgcctg cccgccgggc ttccttatcc
     3841 caccccagag acatcggctg tgaccagcac accagctgcc cggtggggca gacctgctgc
     3901 ccgagcctgg gtgggagctg ggcctgctgc cagttgcccc atgctgtgtg ctgcgaggat
     3961 cgccagcact gctgcccggc tggctacacc tgcaacgtga aggctcgatc ctgcgagaag
     4021 gaagtggtct ctgcccagcc tgccaccttc ctggcccgta gccctcacgt gggtgtgaag
     4081 gacgtggagt gtggggaagg acacttctgc catgataacc agacctgctg ccgagacaac
     4141 cgacagggct gggcctgctg tccctaccgc cagggcgtct gttgtgctga tcggcgccac
     4201 tgctgtcctg ctggcttccg ctgcgcagcc aggggtacca agtgtttgcg cagggaggcc
     4261 ccgcgctggg acgccccttt gagggaccca gccttgagac agctgctggt cgacatgctg
     4321 atccccatcg ctgtgggtgg tgccctggcg gggctggtcc tcatcgtcct catcgcctac
     4381 ctcgtcggca ggaagaggag tcacgcaggc taccagacta tctagtaggt cgacaatcaa
     4441 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4501 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4561 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4621 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4681 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4741 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4801 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     4861 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4921 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4981 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg
     5041 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5101 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     5161 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5221 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5281 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5341 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     5401 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5521 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     5581 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5641 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5701 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     5761 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     5821 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     5881 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     5941 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6001 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6061 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6121 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6181 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6241 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6301 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6361 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6421 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6481 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6541 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6601 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6661 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6721 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6781 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6841 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6901 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6961 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7021 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7081 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7141 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7201 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7261 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7321 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7381 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7441 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7501 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7561 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7621 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7681 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7741 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7801 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7861 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7921 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7981 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8041 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8101 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8161 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8221 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8281 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     8341 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     8401 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     8461 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     8521 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     8581 tgca
//
