LOCUS       sequence-495612-        7153 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Expresses Gluc-feRMA and EGFP under the hSyn promoter. Gluc-feRMA 
            enables monitoring of neuronal transduction and includes a TEV cs, 
            allowing inactivation of it upon interaction with TEV protease..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  AUTHORS   Nouraein S, Lee S, Li H, Saenz VA, Raisely EK, Costa VP, Szablowski 
            JO
  TITLE     Erasable Serum Markers
  JOURNAL   bioRxiv 2025.05.08.652140
REFERENCE   2  (bases 1 to 7153)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.05.08.652140"
COMMENT     Sequence Label: pAAV-hSyn-Gluc-feRMA-IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..7153
                     /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)"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             673..1227
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGEKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIDDIPEIPGFKDLEPIEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGDD"
     CDS             673..1227
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGEKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIDDIPEIPGFKDLEPIEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGDD"
     CDS             1234..1254
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     misc_feature    join(1920..2494,2495..2497,2498..2506)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2087..2104)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2314..2333
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2507..2509,2510..2512,2513..3223)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2507..2509,2510..2512,2513..3223)
                     /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(2552..2573)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2813..2832)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3160..3181
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3233..3253
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3284..3872
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3337..3357)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3755..3766)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3904..4380
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4103..4122)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4420..4560
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4420..4549
                     /label=AAV2 ITR
     rep_origin      4635..5090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4722..4741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4932..4953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5107..5126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5226..5248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5286..5304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5372..5476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5477..5545,5546..6337)
                     /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(5695..5714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6508..7096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6997..7016
                     /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 agtcgagctc
      661 aagcttgcca ccatgggagt caaagttctg tttgccctga tctgcatcgc tgtggccgag
      721 gccaagccca ccgagaacaa cgaagacttc aacatcgtgg ccgtggccag caacttcgcg
      781 accacggatc tcgatgctga ccgcgggaag ttgcccggcg agaagctgcc gctggaggtg
      841 ctcaaagagt tggaagccaa tgcccggaaa gctggctgca ccaggggctg tctgatctgc
      901 ctgtcccaca tcaagtgcac gcccaagatg aagaagttca tcccaggacg ctgccacacc
      961 tacgaaggcg acaaagagtc cgcacagggc ggcataggcg aggcgatcga cgacattcct
     1021 gagattcctg ggttcaagga cttggagccc atagagcagt tcatcgcaca ggtcgatctg
     1081 tgtgtggact gcacaactgg ctgcctcaaa gggcttgcca acgtgcagtg ttctgacctg
     1141 ctcaagaagt ggctgccgca acgctgcgcg acctttgcca gcaagatcca gggccaggtg
     1201 gacaagatca agggggccgg tgatgacggc tccgagaacc tgtacttcca gtccggttca
     1261 gtcccagaag tatcatctgt cttcatcttc cccccaaagc ccaaggatgt gctcaccatt
     1321 actctgactc ctaaggtcac gtgtgttgtg gtagacatca gcaaggatga tcccgaggtc
     1381 cagttcagct ggtttgtaga tgatgtggag gtgcacacag ctcagacgca accccgggag
     1441 gagcagttca acagcacttt ccgctcagtc agtgaacttc ccatcatgca ccaggactgg
     1501 ctcaatggca aggagttcaa atgcagggtc aacagtgcag ctttccctgc ccccatcgag
     1561 aaaaccatct ccaaaaccaa aggcagaccg aaggctccac aggtgtacac cattccacct
     1621 cccaaggagc agatggccaa ggataaagtc agtctgacct gcatgataac agacttcttc
     1681 cctgaagaca ttactgtgga gtggcagtgg aatgggcagc cagcggagaa ctacaagaac
     1741 actcagccca tcatggacac agatggctct tacttcgtct acagcaagct caatgtgcag
     1801 aagagcaact gggaggcagg aaatactttc acctgctctg tgttacatga gggcctgcac
     1861 aaccaccata ctgagaagag cctctcccac tctcctggta aatgattcta ggggatccgc
     1921 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1981 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     2041 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     2101 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     2161 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     2221 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     2281 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     2341 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     2401 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2461 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     2521 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2581 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2641 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2701 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2761 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2821 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2881 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2941 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3001 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3061 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3121 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3181 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtggtt ctcccaagaa
     3241 gaagaggaaa gtctaagttt aattcgatat caagcttatc gataatcaac ctctggatta
     3301 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3361 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3421 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3481 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3541 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3601 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3661 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg
     3721 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3781 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3841 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc
     3901 tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     3961 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     4021 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     4081 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     4141 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     4201 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     4261 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     4321 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     4381 ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     4441 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4501 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4561 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4621 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4681 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4741 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4801 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4861 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4921 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4981 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     5041 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     5101 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5161 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5221 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5281 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5341 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5401 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5461 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5521 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5581 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5641 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5701 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5761 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5821 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5881 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5941 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6001 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6061 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6121 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6181 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6241 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6301 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6361 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6421 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6481 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6541 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6601 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6661 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6721 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6781 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6841 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6901 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6961 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7021 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7081 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7141 gccttttgct cac
//
