LOCUS       sequence-432571-        4939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2431m 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 4939)
  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 4939)
  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: AiP1856 - pAAV-AiE2431m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..85
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(239..258)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     358..380
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(418..436)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        504..608
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(609..677,678..1469)
                     /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(827..846)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1640..2228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2129..2148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2290..2419
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3240..3249
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3246..3248,3249..3251,3252..3962)
                     /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(3291..3312)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3552..3571)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3899..3920
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4055..4075)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4262..4373
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4491..4631
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4491..4620
                     /label=AAV2 ITR
     rep_origin      join(4706..4939,1..222)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4793..4812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
       61 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
      121 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
      181 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
      241 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
      301 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      361 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
      421 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
      481 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      541 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      601 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      661 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      721 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      781 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      841 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      901 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      961 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1021 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1081 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1141 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1201 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1261 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1321 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1381 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1441 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1501 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1561 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1621 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1681 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1741 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1801 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1861 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1921 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1981 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2041 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2101 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2161 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2221 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2281 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
     2341 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     2401 tccatcacta ggggttcctg cggccgcacg cgttgtaaga gtaataatcg ggtaggagtt
     2461 ggggggtggg ggaagaggct aagagagaca gagaaagaca cagatagaga cagacaaaca
     2521 gacagacaga caaacagaga ctgggactag attcggcact tcttgggaac cagtgagaag
     2581 tgcaggcgtg ccatctgagt tccacagatc cccacgggat gtgtttgcac attcacccac
     2641 tctgggggaa gcaggggttc taaggatcaa gaggattgca ggtgactgtg ggattctggg
     2701 aggcctgaat aattgggagg tgggcattca gagatagaac agtgtccctt ggagtccact
     2761 ctgctaccag ggcagctggg ccctggggca cctgccaatc actgcagtgc acttggtaat
     2821 ctcccctctt cacaatcaga tcggtcaagc acccagtgca gctgggcctc tgaattatag
     2881 gctgtgggca gatggctgtc caaggaaaca gccaacgagt gtgcatcagg ccccttcctc
     2941 tggctcctgc tcctgacagc aaacacccac tctctgcaag gaataaggca gtgacacacc
     3001 acatcaaatg cttaccgcat cagagcccac tcccagccct gctcagggaa aaaccctgga
     3061 cctctctgcc ctccaatgca tgattcttgc ctctgacaga tgtaatcctc tcgttcacca
     3121 cacccctccg gtcttagagc taggcggagc tcgggctggg cataaaagtc agggcagagc
     3181 catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg
     3241 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3301 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3361 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     3421 ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca
     3481 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3541 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3601 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3661 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     3721 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     3781 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3841 accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     3901 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3961 agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca
     4021 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     4081 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     4141 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     4201 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg
     4261 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4321 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4381 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4441 tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct
     4501 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4561 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4621 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4681 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4741 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     4801 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4861 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4921 ttgggtgatg gttcacgta
//
