LOCUS       sequence-414343-        6336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV virus production for neuronal expression of dMASS_LF 
            (loss-of-function) under hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6336)
  AUTHORS   Rappleye M, Wait SJ, Lee JD, Siebart JC, Torp L, Smith N, Muster J, 
            Matreyek KA, Fowler DM, Berndt A
  TITLE     Optogenetic Microwell Array Screening System: A High-Throughput 
            Engineering Platform for Genetically Encoded Fluorescent Indicators.
  JOURNAL   ACS Sens. 2023 Nov 24;8(11):4233-4244. doi: 
            10.1021/acssensors.3c01573. Epub 2023 Nov 13.
  PUBMED    37956352
REFERENCE   2  (bases 1 to 6336)
  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"; doi: 
            "10.1021/acssensors.3c01573"; journalName: "ACS Sens"; date: 
            "2023-11-24- 24"; volume: "8"; issue: "11"; pages: "4233-4244"
COMMENT     Sequence Label: pAAV-hsyn-dMASS_LF_WPRE
FEATURES             Location/Qualifiers
     source          1..6336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(75..179)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(205..660)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(342..363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     554..573
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   732..861
                     /label=AAV2 ITR
     promoter        943..1390
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     2412..2433
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2539..2560)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2800..2819)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3505..4093
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3558..3578)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3976..3987)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4096..4112)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4097..4113)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4142..4263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4153..4172)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4207..4226
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4425..4554
                     /label=AAV2 ITR
     rep_origin      complement(4791..5379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4871..4890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5550..6336,1..5,6..74))
                     /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     6173..6192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
       61 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      121 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      181 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
      241 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
      301 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
      361 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
      421 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
      481 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
      541 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
      601 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
      661 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
      721 tattacgcca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
      781 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      841 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      901 agccatgctc taggaagatc tctgcagagg gccctgcgta tgagtgcaag tgggttttag
      961 gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa
     1021 gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag
     1081 gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc
     1141 gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg
     1201 tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc
     1261 cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg
     1321 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct
     1381 gagagcgcag tcgaattcaa gctgctagca aggatcctct agagtcgact ccggagccac
     1441 ctgcgcgcca tgaagacgat catcgccctg agctacatct tctgcctggt gttcgccgac
     1501 tacaaggacg atgatgacgc catgaggact ctgaacacct ctgccatgga cgggactggg
     1561 ctggtggtgg agagggactt ctctgttcgt atcctcactg cctgtttcct gtcgctgctc
     1621 atcctgtcca cgctcctggg gaacacgctg gtctgtgctg ccgttatcag gttccgacac
     1681 ctgcggtcca aggtgaccaa cttctttgtc atctccttgg ctgtgtcaga tctcttggtg
     1741 gccgtcctgg tcatgccctg gaaggcagtg gctgagattg ctggcttctg gccctttggg
     1801 tccttctgta acatctgggt ggcctttgct atcatgtgct ccactgcatc catcctcaac
     1861 ctctgtgtga tcagcgtgga caggtattgg gctatctcca gccctttccg gtatgagaga
     1921 aagatgaccc ccaaggcagc cttcatcctg atcagtgtgg catggacctt gtctgtactc
     1981 atctccttca tcccagtgca gctcagctgg cacaaggcaa aacccacaag cccctctgat
     2041 ggaaatgcca cttccctggc tgagaccata gacaactgtg actccagcct cagcaggaca
     2101 tatgccatct catcctctgt aatcagcttt tacatccctg tggccatcat gattgtcacc
     2161 tacaccagga tctacaggat tgctcagaaa ctgagctcag gttgtaacgt ctatatcaag
     2221 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac
     2281 ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2341 ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga ccccaacgag
     2401 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2461 gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga ggagctgttc
     2521 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2581 gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2641 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2701 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2761 cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2821 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2881 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaaccg gttggaccaa
     2941 ctgaaaagag aaactaaagt cctgaagact ctgtcggtga tcatgggtgt gtttgtgtgc
     3001 tgttggctac ctttcttcat cttgaactgc attttgccct tctgtgggtc tggggagacg
     3061 cagcccttct gcattgattc caacaccttt gacgtgtttg tgtggtttgg gtgggctaat
     3121 tcatccttga accccatcat ttatgccttt aatgctgatt ttcggaaggc attttcaacc
     3181 ctcttaggat gctacagact ttgccctgcg acgaataatg ccatagagac ggtgagtatc
     3241 aataacaatg gggccgcgat gttttccagc catcatgagc cacgaggctc catctccaag
     3301 gagtgcaatc tggtttacct gatcccacat gctgtgggct cctctgagga cctgaaaaag
     3361 gaggaggcag ctggcatcgc cagacccttg gagaagctgt ccccagccct atcggtcata
     3421 ttggactatg acactgacgt ctctctggag aagatccaac ccatcacaca aaacggtcag
     3481 cacccaacct gaaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3541 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3601 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3661 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3721 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3781 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3841 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3901 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3961 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4021 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4081 gccgcctccc cgcatcgata ccgtcgacct cgacccgggc ggccgcttcg agcagacatg
     4141 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4201 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4261 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     4321 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     4381 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     4441 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4501 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat
     4561 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4621 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4681 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4741 aaaggccagc aaaaagccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4801 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4861 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4921 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4981 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5041 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5101 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5161 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5221 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5281 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5341 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5401 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5461 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5521 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5581 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5641 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5701 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5761 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5821 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5881 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5941 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6001 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6061 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6301 tgatcttcag catcttttac tttcaccagc gtttct
//
