LOCUS       sequence-420095-        6391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV virus production for neuonal expression of uMASS 
            (loss-of-function) under hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6391)
  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 6391)
  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_HA_Flag-uMASS_LF
FEATURES             Location/Qualifiers
     source          1..6391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   784..913
                     /label=AAV2 ITR
     polyA_signal    1075..1196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1112..1131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1166..1185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1225..1241
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1226..1242
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1245..1833
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1351..1362
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1760..1780
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     2474..2493
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2733..2754
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2860..2881)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4003..4450)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4532..4661
                     /label=AAV2 ITR
     rep_origin      4733..5188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4820..4839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5030..5051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5214..5318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5319..5387,5388..6179)
                     /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(5537..5556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6350..6391,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gctttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      781 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
      841 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      901 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      961 tctaggaaga tccttatcga ttttaccaca tttgtagagg ttttacttgc tttaaaaaac
     1021 ctcccacatc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1081 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1141 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     1201 gtctgctcga agcggccgcc cgggtcgagg tcgacggtat cgatgcgggg aggcggccca
     1261 aagggagatc cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc gcagagccgg
     1321 cagcaggccg cgggaaggaa ggtccgctgg attgagggcc gaagggacgt agcagaagga
     1381 cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc caaggaaagg acgatgattt
     1441 ccccgacaac accacggaat tgtcagtgcc caacagccga gcccctgtcc agcagcgggc
     1501 aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg gggaaagcga aagtcccgga
     1561 aaggagctga caggtggtgg caatgcccca accagtgggg gttgcgtcag caaacacagt
     1621 gcacaccacg ccacgttgcc tgacaacggg ccacaactcc tcataaagag acagcaacca
     1681 ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg
     1741 cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa
     1801 tctttcacaa attttgtaat ccagaggttg attatcgata agctttcagc cgacttcgtt
     1861 ctcgtagcag aaagcggcgg ctggtgctcc cacgttgatg tcgatctggt ccagagggat
     1921 gtactcgccc tcgcttgtga ttctgctctt acctccgccg ggcaatggag cagtttctgc
     1981 ctccagattt tctagctggt ggttagttcg atccactgta ttagccgtgg agggatgttc
     2041 cctagtgttc tgacggactc gagtggagtt ttgctgttcg atcgtggacg aggttgggat
     2101 gcagaactct ctgaagcatc gcttgaagtt ttcatccagg aaggcgtaaa gaactggatt
     2161 caggcagctg ttcgtgtaac ccaaagcaat gcagaagtgc caggaaacgg tctgaaatgt
     2221 ggtttctgga atcgtgatca gcgctttgat gatgacgtag atgtggatgg gggtccagca
     2281 gacgataaat acagccacga ccaccagcac catccgggtg atcctgcgca gattcctgtc
     2341 cttttctttc agttggtcaa ccatgttgta ctccagcttg tgccccagga tgttgccgtc
     2401 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2461 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggat
     2521 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     2581 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     2641 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcacc
     2701 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     2761 caccaccccg gtgaacagct cctcgccctt gctcaccatg ctccctccgg taccgccctt
     2821 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     2881 gtgatcgcgc ttctcgttgg ggtctttcga aagtttggac tgcacgctca ggtagtggtt
     2941 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtaggcgag
     3001 ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag ttcgccttga tgccgttctt
     3061 ctgcttgtcg gccttgatat agacgttatc aagtgagctc aggcgaacgc tcttgagtcg
     3121 taagatcatc aggccgtaac acacagtgat gatgaggatc ggcatgatga aagcgaagat
     3181 aaagacacag attttgagca ggttctccca gtaccaggtt gggtgggaga acgtgagggt
     3241 gcaatctatg gacccctgcc tgtatttcgt ggttgccatg aacattacag gcagaccgat
     3301 ggcagaagag aggatccagt tgcagacgtt gacgattttg gcatttcggg gggtacggaa
     3361 atccagggct ttgactgggt ggcagacagc aatgtagcgg tccacgctca tggtgcagag
     3421 ggtgaatatg ctggtgaaca tgttgtagta tcctattgag atcacgatct tgcagaggat
     3481 ggttccgaag ggccatgttc ccatcaggta gttgacactc tgaaagggca gtgtactggt
     3541 cgctaaggcg tctgccagag caaggttgaa aatgtagatg ttggtggcag tcttcatttt
     3601 ggtgtatctt acaatcacat acatgaccag gaagtttccg aagaggccca ctacacacac
     3661 gatagagtag agggccatga tggtaatggc tgtgaccatg gaagggctgc cggtctgagg
     3721 gcacaggctg tcgttcccgc caagcccggt gcggttcaga ccgcatggat cggactggtt
     3781 gccatcaacg tgggacaagt tgagccagga gccaggtgct ggggagcaac ttgcctgagc
     3841 taaggggtct gagcagtcgc tggtgttccc tgggccggtg ctgctgtcca tggcgtcatc
     3901 atcgtccttg tagtcggcga acaccaggca gaagatgtag ctcagggcga tgatcgtctt
     3961 catggcgcgc aggtggctcc ggagctagca gcttgaattc gactgcgctc tcaggcacga
     4021 cacgactcct ccgctgccca ccgcagactg aggcagcgct gagtcgccgg cgccgcagcg
     4081 cagatggtcg cgcccgtgcc cccctatctc gcgcctcgcg tggtgcggtc cggctgggcc
     4141 ggcggcggcg cggacgcgac caaggtggcc gggaagggga gtttgcgggg gaccggcgag
     4201 tgacgtcagc gcgccttcag tgctgaggcg gcggtggcgc gcgccgccag gcgggggcga
     4261 aggcactgtc cgcggtgctg aagctggcag tgcgcacgcg cctcgccgca tcctgtttcc
     4321 cctccccctc tctgataggg gatgcgcaat ttggggaatg ggggttgggt gcttgtccag
     4381 tgggtcgggg tcggtcgtca ggtaggcacc cccaccccgc ctcatcctgg tcctaaaacc
     4441 cacttgcact catacgcagg gccctctgca gagatcttcc tagagcatgg ctacgtagat
     4501 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4561 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4621 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa tagcgaagag
     4681 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     4741 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4801 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4861 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4921 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4981 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5041 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5101 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5161 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6361 tttttctgcg cgtaatctgc tgcttgcaaa c
//
