LOCUS       sequence-468256-        8470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing HA-tagged human progranulin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8470)
  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 8470)
  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
FEATURES             Location/Qualifiers
     source          1..8470
                     /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    4321..4909
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4374..4394)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4792..4803)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4996..5229
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5301..5422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5338..5357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5392..5411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5462..5597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5617..5636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5618..5636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5646..5663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5646..5662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5655..5677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5804..6259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5891..5910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6101..6122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6285..6389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6390..6458,6459..7250)
                     /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(6608..6627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7421..8009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7910..7929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8163..8180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8297..8318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8333..8338,8339..8356,8357..8363)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8371..8387
                     /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     8376..8398
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8395..8411
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8395..8411
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8430..8450
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8432..8450
                     /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 agctgctgta atgagtcgac
     4321 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4381 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4441 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4501 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4561 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4621 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4681 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4741 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4801 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4861 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
     4921 gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     4981 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     5041 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5101 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5161 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5221 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     5281 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     5341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5401 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     5461 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5521 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5581 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     5641 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5701 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5761 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     5821 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5881 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5941 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6001 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6061 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6121 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6181 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6241 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6361 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6421 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6481 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6541 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6601 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6661 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6721 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6781 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6841 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6901 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6961 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7021 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7081 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7141 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7201 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7261 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7321 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7381 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7441 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7501 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7561 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7621 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7681 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7741 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7801 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7861 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7921 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7981 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8041 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     8101 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     8161 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     8221 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     8281 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     8341 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     8401 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     8461 tggagctgca
//
