LOCUS       sequence-466334-        4574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2503m 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 4574)
  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 4574)
  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: AiP1974 - pAAV-AiE2503m-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(8..76,77..868)
                     /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(226..245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1039..1627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1528..1547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1689..1818
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2274..2283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2280..2282,2283..2285,2286..2996)
                     /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(2325..2346)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2586..2605)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2933..2954
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3089..3109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3296..3407
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3525..3665
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3525..3654
                     /label=AAV2 ITR
     rep_origin      3740..4195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3827..3846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4037..4058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4212..4231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4331..4353
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4391..4409)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4477..4574,1..7)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
       61 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      121 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      181 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      241 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      301 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      361 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      421 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      481 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      541 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      601 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      661 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      721 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      781 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      841 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      901 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      961 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1021 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1081 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1141 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1501 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1561 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1621 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1681 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
     1741 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     1801 ccatcactag gggttcctgc ggccgcacgc gttgtaagag taataatcgg cctgtgccct
     1861 ggaggggact tgaagacatc caaaaagcta gtgagagggc tcctagattt atttgtctcc
     1921 aagggctata tatagccttc ctaacatgaa cccttgggta atccagcatg ggcgctccca
     1981 tatgccctgg tttgattaga gagctctaga tgtctcctgt cccagaacac cagccagccc
     2041 ctgtcttcat gtcgtgtcta gggcggaggg tgattcagag gcaggtgcct gcgacagtgg
     2101 atgcaattag atctaatggg acggaggcct ctctcgtccg tcgccctcgc tctgtgccca
     2161 cccccgcctc cctcaggcac agcaggcgtg gagaggatgc gcaggaggta ggaggtgggg
     2221 gacccagagg ggctttgacg tcagcctggc ctttaagagg cccgctaccg gtcgccacca
     2281 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2341 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2401 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2461 tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
     2521 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2581 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2641 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2701 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     2761 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     2821 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2881 acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2941 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     3001 tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt
     3061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3181 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3241 ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg
     3301 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3361 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3421 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3481 gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3541 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3601 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3661 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3721 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3781 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     3841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     3961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4081 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4141 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4201 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4261 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4321 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4381 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4441 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4501 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4561 taatattgaa aaag
//
