LOCUS       sequence-501099-        8594 bp    DNA     circular SYN 25-FEB-2026
DEFINITION  Lentiviral expression of EGFP-mouse Parkin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8594)
  TITLE     David Chan Lab Plasmids Jan 2026
REFERENCE   2  (bases 1 to 8594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 25, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pHAGE EGFP-mouse Parkin
FEATURES             Location/Qualifiers
     source          1..8594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2062..2179
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2231..2534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2535..2738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2689..2709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2735..2759
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2828..2830,2831..2833,2834..3544)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2828..2830,2831..2833,2834..3544)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2873..2894)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3134..3153)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3481..3502
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4984..5572
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5037..5057)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5455..5466)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5647..5880
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5999..6017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6085..6189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6190..6258,6259..7050)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6408..6427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7221..7809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7710..7729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7963..7980
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8220..8241
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8256..8261,8262..8279,8280..8286)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8294..8310
                     /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     8299..8321
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8318..8334
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8318..8334
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8384..8403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttaattcaca
     2041 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     2101 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     2161 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggactag
     2221 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2281 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2341 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2401 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2461 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2521 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2581 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2641 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2701 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     2761 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccggcgg ccgcggtacc
     2821 taaaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2881 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2941 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3001 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3061 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3121 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3181 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3241 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3301 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3361 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3421 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3481 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3541 caagtccggc cggactcaga tcattggatc aattccaatg atagtgtttg tcaggttcaa
     3601 ctccagctat ggcttcccag tggaggtcga ttctgacacc agcatcttgc agctcaagga
     3661 agtggttgct aagcgacagg gggttccagc tgaccagctg cgtgtgattt ttgccgggaa
     3721 ggagcttccg aatcacctga cggttcaaaa ctgtgacctg gaacaacaga gtattgtaca
     3781 catagtacag agaccacgga ggagaagtca tgaaacaaat gcatctggag gggacgaacc
     3841 ccagagcacc tcagagggct ccatatggga gtccaggagc ttgacacgag tggacctgag
     3901 cagccatacc ctgccggtgg actctgtggg gctggcggtc attctggaca cagacagtaa
     3961 gagggattca gaagcagcca gaggtccagt taaacccacc tacaacagct ttttcatcta
     4021 ctgcaaaggc ccctgccaca aggtccagcc tggaaagctc cgagttcagt gtggcacctg
     4081 caaacaagca accctcacct tggcccaggg cccatcttgc tgggacgatg tcttaattcc
     4141 aaaccggatg agtggtgagt gccagtctcc agactgccct ggaaccagag ctgaattttt
     4201 ctttaaatgt ggagcacacc caacctcaga caaggacacg tcggtagctt tgaacctgat
     4261 caccagcaac aggcgcagca tcccttgcat agcgtgcaca gatgtcagga gccctgtcct
     4321 ggtcttccag tgtaaccacc gtcacgtgat ctgtttggac tgtttccact tgtattgtgt
     4381 cacaagactc aacgatcggc agtttgtcca cgatgctcaa cttggctact ccctgccgtg
     4441 tgtagctggc tgtcccaact ccctgattaa agagctccat cacttcagga tccttggaga
     4501 agagcagtac actaggtacc agcagtatgg ggccgaggaa tgcgtgctgc aaatgggagg
     4561 tgtgctgtgc ccccgtcctg gctgtggagc tggactgcta cctgaacagg gccagaggaa
     4621 agtcacctgc gaagggggca acggcctggg ctgcgggttt gttttctgcc gggactgtaa
     4681 ggaagcatac catgaagggg attgcgactc actgctcgaa ccctcaggag ccacttctca
     4741 ggcctacagg gtggacaaaa gagccgctga gcaagctcgc tgggaggagg cctccaagga
     4801 aaccatcaag aagaccacca agccttgtcc tcgctgcaac gtgccaattg aaaaaaacgg
     4861 aggatgtatg cacatgaagt gtcctcagcc ccagtgcaag ctggagtggt gctggaactg
     4921 tggctgtgag tggaaccgag cctgcatggg agatcactgg tttgacgtgt agctcgagat
     4981 cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5041 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5101 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5161 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5221 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5281 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5341 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     5401 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5461 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5521 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgagatc
     5581 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     5641 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     5701 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     5761 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     5821 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     5881 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     5941 agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc
     6001 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6421 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6781 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6901 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7021 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7861 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7921 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7981 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8041 tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     8101 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     8161 agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8221 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8281 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt
     8341 acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8401 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     8461 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     8521 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     8581 acaaacttag tagt
//
