LOCUS       sequence-376526-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gluc-RMA and EGFP under the hSyn promoter. Gluc-RMA for 
            monitoring neuronal transduction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  AUTHORS   Lee S, Nouraein S, Kwon JJ, Huang Z, Wojick JA, Xia B, Corder G, 
            Szablowski JO
  TITLE     Engineered serum markers for non-invasive monitoring of gene 
            expression in the brain.
  JOURNAL   Nat Biotechnol. 2024 Jan 10. doi: 10.1038/s41587-023-02087-x.
  PUBMED    38200117
REFERENCE   2  (bases 1 to 7126)
  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: "Nat 
            Biotechnol. 2024 Jan 10. doi: 10.1038/s41587-023-02087-x."
COMMENT     Sequence Label: pAAV-hSyn-Gluc-RMA-IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   21..150
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        195..642
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      683..692
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             689..1243
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGEKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIDDIPEIPGFKDLEPIEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGDD"
     CDS             689..1243
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGEKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIDDIPEIPGFKDLEPIEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGDD"
     misc_feature    join(1909..2483,2484..2486,2487..2495)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2076..2093)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2303..2322
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2496..2498,2499..2501,2502..3212)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2496..2498,2499..2501,2502..3212)
                     /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(2541..2562)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2802..2821)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3149..3170
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3222..3242
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3273..3861
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3326..3346)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3744..3755)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3893..4369
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4092..4111)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4409..4549
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4409..4538
                     /label=AAV2 ITR
     rep_origin      4624..5079
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4711..4730)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4921..4942
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5096..5115)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5215..5237
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5275..5293)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5361..5465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5466..5534,5535..6326)
                     /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(5684..5703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6497..7085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6986..7005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
       61 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      121 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtgtgtct agactgcaga
      181 gggccctgcg tatgagtgca agtgggtttt aggaccagga tgaggcgggg tgggggtgcc
      241 tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg
      301 catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca
      361 gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc
      421 agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac
      481 cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata
      541 ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct
      601 gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc
      661 ctctagagtc gagctcaagc ttgccaccat gggagtcaaa gttctgtttg ccctgatctg
      721 catcgctgtg gccgaggcca agcccaccga gaacaacgaa gacttcaaca tcgtggccgt
      781 ggccagcaac ttcgcgacca cggatctcga tgctgaccgc gggaagttgc ccggcgagaa
      841 gctgccgctg gaggtgctca aagagttgga agccaatgcc cggaaagctg gctgcaccag
      901 gggctgtctg atctgcctgt cccacatcaa gtgcacgccc aagatgaaga agttcatccc
      961 aggacgctgc cacacctacg aaggcgacaa agagtccgca cagggcggca taggcgaggc
     1021 gatcgacgac attcctgaga ttcctgggtt caaggacttg gagcccatag agcagttcat
     1081 cgcacaggtc gatctgtgtg tggactgcac aactggctgc ctcaaagggc ttgccaacgt
     1141 gcagtgttct gacctgctca agaagtggct gccgcaacgc tgcgcgacct ttgccagcaa
     1201 gatccagggc caggtggaca agatcaaggg ggccggtgat gacggctccg tcccagaagt
     1261 atcatctgtc ttcatcttcc ccccaaagcc caaggatgtg ctcaccatta ctctgactcc
     1321 taaggtcacg tgtgttgtgg tagacatcag caaggatgat cccgaggtcc agttcagctg
     1381 gtttgtagat gatgtggagg tgcacacagc tcagacgcaa ccccgggagg agcagttcaa
     1441 cagcactttc cgctcagtca gtgaacttcc catcatgcac caggactggc tcaatggcaa
     1501 ggagttcaaa tgcagggtca acagtgcagc tttccctgcc cccatcgaga aaaccatctc
     1561 caaaaccaaa ggcagaccga aggctccaca ggtgtacacc attccacctc ccaaggagca
     1621 gatggccaag gataaagtca gtctgacctg catgataaca gacttcttcc ctgaagacat
     1681 tactgtggag tggcagtgga atgggcagcc agcggagaac tacaagaaca ctcagcccat
     1741 catggacaca gatggctctt acttcgtcta cagcaagctc aatgtgcaga agagcaactg
     1801 ggaggcagga aatactttca cctgctctgt gttacatgag ggcctgcaca accaccatac
     1861 tgagaagagc ctctcccact ctcctggtaa atgattctag gggatccgcc cctctccctc
     1921 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1981 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     2041 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     2101 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     2161 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     2221 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     2281 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     2341 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca catgctttac
     2401 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     2461 cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc gaggagctgt
     2521 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2581 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2641 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2701 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2761 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2821 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2881 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2941 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     3001 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3061 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     3121 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3181 ggatcactct cggcatggac gagctgtaca agggtggttc tcccaagaag aagaggaaag
     3241 tctaagttta attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     3301 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3361 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3421 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3481 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3541 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3601 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3661 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     3721 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3781 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3841 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     3901 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     3961 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     4021 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     4081 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     4141 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     4201 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     4261 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4321 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     4381 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4441 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4501 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4561 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4621 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4681 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4741 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4801 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4861 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4921 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4981 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     5041 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5101 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5161 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5221 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5281 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5341 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5401 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5461 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5521 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5581 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5641 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5701 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5761 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5821 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5881 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5941 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6001 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6061 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6121 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6181 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6241 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6301 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6361 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6421 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6481 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6541 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6601 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6661 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6721 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6781 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6841 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6901 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6961 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7021 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7081 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttg
//
