LOCUS       sequence-450599-        6918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cluc-NHP.RMA under the hSyn promoter. Cluc-NHP.RMA for 
            monitoring neuronal transduction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6918)
  AUTHORS   Lee S, Romac M, Watanabe S, Chernov M, Li H, Raisley E, Szablowski 
            HO, Costa VD
  TITLE     Synthetic Serum Markers Enable Noninvasive Monitoring of Gene 
            Expression in Primate Brains
  JOURNAL   bioRxiv 2025.06.01.657212; doi: 10.1101/2025.06.01.657212
REFERENCE   2  (bases 1 to 6918)
  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: "bioRxiv 
            2025.06.01.657212; doi: 10.1101/2025.06.01.657212"
COMMENT     Sequence Label: pAAV-hSyn-Cluc-NHP.RMA
FEATURES             Location/Qualifiers
     source          1..6918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..134
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        179..626
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             673..2331
                     /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"
     CDS             2986..3018
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     misc_feature    3049..3637
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3102..3122)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3520..3531)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3669..4145
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3868..3887)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4185..4325
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4185..4314
                     /label=AAV2 ITR
     rep_origin      4400..4855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4487..4506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4697..4718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4872..4891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4991..5013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5051..5069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5137..5241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5242..5310,5311..6102)
                     /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(5460..5479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6273..6861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6762..6781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
       61 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      121 cactaggggt tcctgcggcc gcacgcgtgt gtctagactg cagagggccc tgcgtatgag
      181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
      241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
      301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
      361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
      421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
      481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
      541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      601 gagtcgtgtc gtgcctgaga gcgcagtcga gaaggtaccg gatcctctag agtcgagcta
      661 aagcttgcca ccatgaagac cttaattctt gccgttgcat tagtctactg cgccactgtt
      721 cattgccagg actgtcctta cgaacctgat ccaccaaaca cagttccaac ttcctgtgaa
      781 gctaaagaag gagaatgtat tgatagcagc tgtggcacct gcacgagaga catactatca
      841 gatggactgt gtgaaaataa accaggaaaa acatgttgcc gaatgtgtca gtatgtaatt
      901 gaatgcagag tagaggccgc aggatggttt agaacattct atggaaagag attccagttc
      961 caggaacctg gtacatacgt gttgggtcaa ggaaccaagg gcggcgactg gaaggtgtcc
     1021 atcaccctgg agaacctgga tggaaccaag ggggctgtgc tgaccaagac aagactggaa
     1081 gtggctggag acatcattga catcgctcaa gctactgaga atcccatcac tgtaaacggt
     1141 ggagctgacc ctatcatcgc caacccgtac accatcggcg aggtcaccat cgctgttgtt
     1201 gagatgccag gcttcaacat caccgtcatt gagttcttca aactgatcgt gatcgacatc
     1261 ctcggaggaa gatctgtaag aatcgcccca gacacagcaa acaaaggaat gatctctggc
     1321 ctctgtggag atcttaaaat gatggaagat acagacttca cttcagatcc agaacaactc
     1381 gctattcagc ctaagatcaa ccaggagttt gacggttgtc cactctatgg aaatcctgat
     1441 gacgttgcat actgcaaagg tcttctggag ccgtacaagg acagctgccg caaccccatc
     1501 aacttctact actacaccat ctcctgcgcc ttcgcccgct gtatgggtgg agacgagcga
     1561 gcctcacacg tgctgcttga ctacagggag acgtgcgctg ctcccgaaac tagaggaacc
     1621 tgcgttttgt ctggacatac tttctacgat acatttgaca aagcaagata ccaattccag
     1681 ggtccctgca aggagattct tatggccgcc gactgtttct ggaacacttg ggatgtgaag
     1741 gtttcacaca ggaatgttga ctcttacact gaagtagaga aagtacgaat caggaaacaa
     1801 tcgactgtag tagaactcat tgttgatgga aaacagattc tggttggagg agaagccgtg
     1861 tccgtcccgt acagctctca gaacacttcc atctactggc aagatggtga catactgact
     1921 acagccatcc tacctgaagc tctggtggtc aagttcaact tcaagcaact gctcgtcgta
     1981 catattagag atccattcga tggtaagact tgcggtattt gcggtaacta caaccaggat
     2041 ttcagtgatg attcttttga tgctgaagga gcctgtgatc tgacccccaa cccaccggga
     2101 tgcaccgaag aacagaaacc tgaagctgaa cgactctgca atagtctctt cgccggtcaa
     2161 agtgatcttg atcagaaatg taacgtgtgc cacaagcctg accgtgtcga acgatgcatg
     2221 tacgagtatt gcctgagggg acaacagggt ttctgtgacc acgcatggga gttcaagaaa
     2281 gaatgctaca taaagcatgg agacacccta gaagtaccag atgaatgcaa aggctccctc
     2341 ctggggggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc
     2401 cggacccctg aggtcacatg cgtggtggta gacgtgagcc aggaagaccc cgatgtcaag
     2461 ttcaactggt acgtaaacgg cgcggaggtg catcatgccc agacgaagcc acgggagacg
     2521 cagtacaaca gcacatatcg tgtggtcagc gtcctcaccg tcacgcacca ggactggctg
     2581 aacggcaagg agtacacgtg caaggtctcc aacaaagccc tcccggcccc catccagaaa
     2641 accatctcca aagacaaagg gcagccccga gagcctcagg tgtacaccct gcccccgtcc
     2701 cgggaggagc tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg cttctacccc
     2761 agcgacatcg tcgtggagtg ggagagcagc gggcagccgg agaacaccta caagaccacg
     2821 ccgcccgtgc tggactccga cggctcctac ttcctctaca gcaagctcac cgtggacaag
     2881 agcaggtggc agcaggggaa cgtcttctca tgctccgtga tgcatgaggc tctgcacaac
     2941 cactacaccc agaagagcct ctccgtgtct ccgggtaaag gttctatggc tagcatgact
     3001 ggtggacagc aaatgggttg agtttaattc gatatcaagc ttatcgataa tcaacctctg
     3061 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3121 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3181 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3241 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3301 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3361 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3421 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3481 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3541 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3601 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3661 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3721 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3781 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3841 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3901 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3961 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     4021 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     4081 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     4141 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4201 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4261 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4321 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4381 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4441 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4501 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4561 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4621 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4681 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4741 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4801 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4861 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4921 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4981 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5041 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5101 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5161 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5221 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5281 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5341 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5401 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5461 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5521 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5581 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5641 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5701 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5761 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5821 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5881 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5941 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6001 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6061 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6121 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6181 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6241 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6301 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6361 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6421 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6481 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6541 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6601 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6661 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6721 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6781 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6841 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6901 tgctggcctt ttgctcac
//
