LOCUS       sequence-414406-        6391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV virus production for neuronal expression of uMASS_1 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_1_WPRE
FEATURES             Location/Qualifiers
     source          1..6391
                     /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     2512..2533
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2639..2660)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2900..2919)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3560..4148
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3613..3633)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4031..4042)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4151..4167)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4152..4168)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4197..4318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4208..4227)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4262..4281
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4480..4609
                     /label=AAV2 ITR
     rep_origin      complement(4846..5434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4926..4945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5605..6391,1..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     6228..6247
                     /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 gctgctagct ccggagccac ctgcgcgcca tgaagacgat
     1441 catcgccctg agctacatct tctgcctggt gttcgccgac tacaaggacg atgatgacgc
     1501 catggacagc agcaccggcc cagggaacac cagcgactgc tcagacccct tagctcaggc
     1561 aagttgctcc ccagcacctg gctcctggct caacttgtcc cacgttgatg gcaaccagtc
     1621 cgatccatgc ggtctgaacc gcaccgggct tggcgggaac gacagcctgt gccctcagac
     1681 cggcagccct tccatggtca cagccattac catcatggcc ctctactcta tcgtgtgtgt
     1741 agtgggcctc ttcggaaact tcctggtcat gtatgtgatt gtaagataca ccaaaatgaa
     1801 gactgccacc aacatctaca ttttcaacct tgctctggca gacgccttag cgaccagtac
     1861 actgcccttt cagagtgtca actacctgat gggaacatgg cccttcggaa ccatcctctg
     1921 caagatcgtg atctcaatag attactacaa catgttcacc agcatattca ccctctgcac
     1981 catgagcgtg gaccgctaca ttgctgtctg ccacccagtc aaagccctgg atttccgtac
     2041 cccccgaaat gccaaaatcg tcaacgtctg caactggatc ctctcttctg ccatcggtct
     2101 gcctgtaatg ttcatggcaa ccacgaaata caggcagggg tccatagatt gcaccctcac
     2161 gttctcccac ccaacctggt actgggagaa cctgctcaaa atctgtgtct ttatcttcgc
     2221 tttcatcatg ccgatcctca tcatcactgt gtgttacggc ctgatgatct tacgactcaa
     2281 gagcgttcgc ctgagctcac ttgataacgt ctatatcaag gccgacaagc agaagaacgg
     2341 catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccta
     2401 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2461 cctgagcgtg cagtccaaac tttcgaaaga ccccaacgag aagcgcgatc acatggtcct
     2521 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcgg
     2581 taccggaggg agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2641 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggtgaggg
     2701 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2761 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2821 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acatccagga
     2881 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2941 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3001 catcctgggg cacaagctgg agtacaacat ggttgaccaa ctgaaagaaa aggacaggaa
     3061 tctgcgcagg atcacccgga tggtgctggt ggtcgtggct gtatttatcg tctgctggac
     3121 ccccatccac atctacgtca tcatcaaagc gctgatcacg attccagaaa ccacatttca
     3181 gaccgtttcc tggcacttct gcattgcttt gggttacacg aacagctgcc tgaatccagt
     3241 tctttacgcc ttcctggatg aaaacttcaa gcgatgcttc agagagttct gcatcccaac
     3301 ctcgtccacg atcgaacagc aaaactccac tcgagtccgt cagaacacta gggaacatcc
     3361 ctccacggct aatacagtgg atcgaactaa ccaccagcta gaaaatctgg aggcagaaac
     3421 tgctccattg cccggcggag gtaagagcag aatcacaagc gagggcgagt acatccctct
     3481 ggaccagatc gacatcaacg tgggagcacc agccgccgct ttctgctacg agaacgaagt
     3541 cggctgaaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3601 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3661 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3721 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3781 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3841 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3901 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3961 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     4021 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4081 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4141 ctccccgcat cgataccgtc gacctcgacc cgggcggccg cttcgagcag acatgataag
     4201 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4261 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4321 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     4381 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     4441 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     4501 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4561 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg
     4621 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4681 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4741 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4801 ccagcaaaaa gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4861 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4921 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4981 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5041 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5101 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5161 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5221 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5281 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5341 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5401 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5461 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5521 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5581 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5641 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5701 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5761 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5821 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5881 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5941 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6001 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6061 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6121 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6181 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6241 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6301 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6361 ttcagcatct tttactttca ccagcgtttc t
//
