LOCUS       sequence-473581-        4989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0377m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4989)
  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: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP13060 - pAAV-AiE0377m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1001..1010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1007..1009,1010..1012,1013..1723)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1052..1073)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1313..1332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1660..1681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1816..1836)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2023..2134
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2252..2392
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2252..2381
                     /label=AAV2 ITR
     rep_origin      2467..2922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2554..2573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2764..2785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2939..2958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3058..3080
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3118..3136)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3204..3308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3309..3377,3378..4169)
                     /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(3527..3546)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4340..4928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4829..4848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagcctg gtcctggaat agcacccaga tatactgggg
      181 aggttcatga aagagaaaag gtctaagctg agagtggtca ttagcctgag agatctgacc
      241 tttcacagaa aagatttagt gtaattcggt accgcgtgag tgtgcttata aaatgccaat
      301 tcacctccac tctcagaaga cttttctaac atgcacctca tttcttacta tgtatctggc
      361 ctgaaatagg agcattggct aaaggtgagg attcccaggg gttgctactg taattaaaac
      421 agaaagccag ttcttggaga catggacact cagaacttga tggttaatga ttatggttaa
      481 tgccatctgg ttatgagcag tagaattcta gggtggggga tactaagagt ctgctgtaat
      541 atacatacta ctcaaatgta agatgatatg cttatattta tgatccttta acatcccaat
      601 ctctggacct cagaagagaa gactggctct gggagctaat taacatgcag tcaatatccc
      661 aagtctctct ttgttcaaga aggcacaggc tgaactcatt aaactgccag cagaccagaa
      721 aaataagtta tttgaaagcc aacacaggaa aacttattta taaagctcag cagattttaa
      781 aagatcaaaa caaaacccat taataatagg ctgggcttgt agctaagtag agtgattgcg
      841 tgaagtcttg ggttcaggtt cgattcccag cgcagggtca actttgtaag gcagtacacg
      901 cttgcaagag ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg
      961 cttctgggat ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg
     1021 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1081 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1141 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1201 gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1261 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1321 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1381 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1441 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     1501 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     1561 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     1621 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1681 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc
     1741 gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac
     1801 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1861 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1921 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1981 gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc
     2041 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2101 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2161 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2221 atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     2281 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2341 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     2401 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2461 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2521 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     2581 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2641 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     2701 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2761 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     2821 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     2881 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     2941 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3001 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3061 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3121 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3181 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3241 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3301 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3361 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3421 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3481 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3541 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3601 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3661 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3721 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3781 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3841 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3901 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3961 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4021 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4081 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4141 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4201 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4261 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4321 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4381 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4441 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4501 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4561 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4621 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4681 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4741 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4801 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4861 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4921 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4981 ctcacatgt
//
