LOCUS       sequence-376506-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cluc-RMA under the hSyn promoter. Cluc-RMA for monitoring 
            neuronal transduction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  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 6879)
  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-Cluc-RMA
FEATURES             Location/Qualifiers
     source          1..6879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        209..656
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             703..2361
                     /codon_start=1
                     /product="secreted Cypridina luciferase"
                     /label=CLuc
                     /note="contains 30 codon substitutions for efficient 
                     translation in mammalian cells"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     misc_feature    3040..3628
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3093..3113)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3511..3522)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3660..4136
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3859..3878)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4176..4316
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4176..4305
                     /label=AAV2 ITR
     rep_origin      4391..4846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4478..4497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4688..4709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4863..4882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4982..5004
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5042..5060)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5128..5232
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5233..5301,5302..6093)
                     /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(5451..5470)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6264..6852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6753..6772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg
       61 ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgtgt
      181 gtctagactg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
      241 ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca
      301 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
      361 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
      421 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
      481 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
      541 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
      601 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
      661 gaaggtaccg gatcctctag agtcgagcta aagcttgcca ccatgaagac cttaattctt
      721 gccgttgcat tagtctactg cgccactgtt cattgccagg actgtcctta cgaacctgat
      781 ccaccaaaca cagttccaac ttcctgtgaa gctaaagaag gagaatgtat tgatagcagc
      841 tgtggcacct gcacgagaga catactatca gatggactgt gtgaaaataa accaggaaaa
      901 acatgttgcc gaatgtgtca gtatgtaatt gaatgcagag tagaggccgc aggatggttt
      961 agaacattct atggaaagag attccagttc caggaacctg gtacatacgt gttgggtcaa
     1021 ggaaccaagg gcggcgactg gaaggtgtcc atcaccctgg agaacctgga tggaaccaag
     1081 ggggctgtgc tgaccaagac aagactggaa gtggctggag acatcattga catcgctcaa
     1141 gctactgaga atcccatcac tgtaaacggt ggagctgacc ctatcatcgc caacccgtac
     1201 accatcggcg aggtcaccat cgctgttgtt gagatgccag gcttcaacat caccgtcatt
     1261 gagttcttca aactgatcgt gatcgacatc ctcggaggaa gatctgtaag aatcgcccca
     1321 gacacagcaa acaaaggaat gatctctggc ctctgtggag atcttaaaat gatggaagat
     1381 acagacttca cttcagatcc agaacaactc gctattcagc ctaagatcaa ccaggagttt
     1441 gacggttgtc cactctatgg aaatcctgat gacgttgcat actgcaaagg tcttctggag
     1501 ccgtacaagg acagctgccg caaccccatc aacttctact actacaccat ctcctgcgcc
     1561 ttcgcccgct gtatgggtgg agacgagcga gcctcacacg tgctgcttga ctacagggag
     1621 acgtgcgctg ctcccgaaac tagaggaacc tgcgttttgt ctggacatac tttctacgat
     1681 acatttgaca aagcaagata ccaattccag ggtccctgca aggagattct tatggccgcc
     1741 gactgtttct ggaacacttg ggatgtgaag gtttcacaca ggaatgttga ctcttacact
     1801 gaagtagaga aagtacgaat caggaaacaa tcgactgtag tagaactcat tgttgatgga
     1861 aaacagattc tggttggagg agaagccgtg tccgtcccgt acagctctca gaacacttcc
     1921 atctactggc aagatggtga catactgact acagccatcc tacctgaagc tctggtggtc
     1981 aagttcaact tcaagcaact gctcgtcgta catattagag atccattcga tggtaagact
     2041 tgcggtattt gcggtaacta caaccaggat ttcagtgatg attcttttga tgctgaagga
     2101 gcctgtgatc tgacccccaa cccaccggga tgcaccgaag aacagaaacc tgaagctgaa
     2161 cgactctgca atagtctctt cgccggtcaa agtgatcttg atcagaaatg taacgtgtgc
     2221 cacaagcctg accgtgtcga acgatgcatg tacgagtatt gcctgagggg acaacagggt
     2281 ttctgtgacc acgcatggga gttcaagaaa gaatgctaca taaagcatgg agacacccta
     2341 gaagtaccag atgaatgcaa aggctccgtc ccagaagtat catctgtctt catcttcccc
     2401 ccaaagccca aggatgtgct caccattact ctgactccta aggtcacgtg tgttgtggta
     2461 gacatcagca aggatgatcc cgaggtccag ttcagctggt ttgtagatga tgtggaggtg
     2521 cacacagctc agacgcaacc ccgggaggag cagttcaaca gcactttccg ctcagtcagt
     2581 gaacttccca tcatgcacca ggactggctc aatggcaagg agttcaaatg cagggtcaac
     2641 agtgcagctt tccctgcccc catcgagaaa accatctcca aaaccaaagg cagaccgaag
     2701 gctccacagg tgtacaccat tccacctccc aaggagcaga tggccaagga taaagtcagt
     2761 ctgacctgca tgataacaga cttcttccct gaagacatta ctgtggagtg gcagtggaat
     2821 gggcagccag cggagaacta caagaacact cagcccatca tggacacaga tggctcttac
     2881 ttcgtctaca gcaagctcaa tgtgcagaag agcaactggg aggcaggaaa tactttcacc
     2941 tgctctgtgt tacatgaggg cctgcacaac caccatactg agaagagcct ctcccactct
     3001 cctggtaaat gagtttaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     3061 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3121 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3181 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3241 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3301 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3361 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3421 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     3481 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3541 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3601 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg
     3661 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     3721 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     3781 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     3841 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     3901 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     3961 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     4021 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     4081 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     4141 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4201 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4261 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4321 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4381 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4441 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     4501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4681 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4741 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4801 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4861 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4921 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4981 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5041 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5101 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5161 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5221 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5281 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5341 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5401 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5461 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5521 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5581 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5641 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5701 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5761 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5821 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5881 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5941 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6001 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6061 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6121 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6181 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6241 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6301 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6361 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6421 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6481 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6541 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6601 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6661 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6721 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6781 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6841 agcctatgga aaaacgccag caacgcggcc tttttacgg
//
