LOCUS       sequence-466197-        4674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3053m 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 4674)
  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 4674)
  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: AiP20043 - pAAV-AiE3053m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      231..240
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(237..239,240..242,243..953)
                     /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(282..303)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(543..562)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     890..911
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1046..1066)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1253..1364
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1482..1622
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1482..1611
                     /label=AAV2 ITR
     rep_origin      1697..2152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1784..1803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1994..2015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2169..2188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2288..2310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2348..2366)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2434..2538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2539..2607,2608..3399)
                     /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(2757..2776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3570..4158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4059..4078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4220..4349
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tacgtatcct gaatcacagt ccactggggg gaagccaagg caggaactca aatcaggaag
       61 gagcctggag gcaggagctg ctgcagagcc cacgggagag tgctgcttgc tgccttgctt
      121 cccatggctc gccggcggag ctcgggctgg gcataaaagt cagggcagag ccatctattg
      181 cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc gccaccatgg
      241 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      301 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      361 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      421 tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
      481 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      541 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      601 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
      661 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
      721 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
      781 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
      841 tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
      901 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg
      961 acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg
     1021 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1081 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1141 aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1201 caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct
     1261 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1321 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1381 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1441 aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     1501 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     1561 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     1621 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     1681 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1741 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     1801 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1861 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     1921 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1981 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     2041 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     2101 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     2161 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     2221 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2281 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     2341 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2401 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2461 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2521 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2581 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2641 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2701 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2761 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2821 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2881 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2941 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3001 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3061 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3121 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3181 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3241 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3301 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3361 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3421 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3481 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3541 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3601 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3661 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3721 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3781 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3841 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3901 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3961 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4021 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4081 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4141 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4201 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     4261 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     4321 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtaaaaaaa tggcatttta
     4381 tccaaatctg gtattcaaaa actgttcatg tcacacactc tgactcagcc aagcttgtgt
     4441 tacctcattg ctgaatttcc ctagaatgtt tcatcagtga ttgtcttctg tgagtattat
     4501 ggttggaagg aggaattgca ctcagcccgc ctgcttgctt gcagctttat attagagtcc
     4561 tggattatat tccacctaac gagtccagta tgtagagata ttgctttaga tagggtttct
     4621 tttgctgtga taaaatcacc atggcaactt gggaaggaaa gggtttcctt ggtt
//
