LOCUS       sequence-474058-        5209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0122h 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 5209)
  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 5209)
  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: AiP11568 -
            pAAV-hsA2-AiE0122h-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5209
                     /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      1246..1255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1252..1254,1255..1257,1258..1968)
                     /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(1297..1318)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1558..1577)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1905..1926
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2036..2056)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2243..2354
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2472..2612
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2472..2601
                     /label=AAV2 ITR
     rep_origin      2687..3142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2774..2793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2984..3005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3159..3178)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3278..3300
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3338..3356)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3424..3528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3529..3597,3598..4389)
                     /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(3747..3766)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4560..5148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5049..5068
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttctg
      361 gaatgaggac acttgttgtg caccttgctt tggagcccca gacaggcaaa aaaatatcca
      421 tgtggcttca cccaggtcat ctctttgctt gtatgaacct caggccaccc ttagaaaggg
      481 tctcatagcc ccggtgtctc actctctgac ctttgccaag tcgtcttcag cagcagctgg
      541 aactctttga agtggctgcc acagatgttg gcaaagcccg gcctgggatg cctccaccct
      601 gcaccaggca cggggctgcc tctgctgcat attcacctgg aaatcatcca catgccaagc
      661 accgcagagg caaggggagt ctgccattgg actgcgttca acacggctgc agggatacag
      721 acaagcacat tccctgtcag aggcacgggc ccggctggct ggagctgcag cagctgtgtg
      781 tggtcacaga gcttccaaat gagggagatg gatggcctcg cttcacagtg tgcccagagc
      841 atgtccacgg tgcctgggaa gaccagggac tcaccatgcc cggctctggt ttccctcaca
      901 gcgaagcctc cttgctggta ggatccatgg ccaagcatgg gggcaggggc tgggggagct
      961 ctgtcagtga cttctcgcag gcagcaaggc ctctgggagc tggccaggaa aacagaaact
     1021 ccatcctgga gcaggtgcgc aaaccaacag ctcgaattcg atatcataat caaccatagg
     1081 taccgagctc gggattcagc cgggagctta gggaggggag gtcacttcat aagggcctgg
     1141 ggggggagtt ggagccacga gtcgtccagc cggagccccg tgtggctgag ctccggcctc
     1201 agaagcatcc ccgggttgga tccttcgaag ctagcgctac cggtcgccac catggtgagc
     1261 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1321 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1381 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1441 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1501 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1561 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1621 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1681 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1741 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1801 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1861 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1921 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacatc
     1981 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2041 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2101 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     2161 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2221 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2281 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2341 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2401 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     2461 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2521 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     2581 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     2641 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     2701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2761 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2881 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3001 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     3061 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgg gaattttaac
     3121 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     3181 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3241 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3301 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3361 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3481 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3541 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3601 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3661 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3721 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3781 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3841 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3901 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3961 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4021 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4081 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4141 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4201 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4261 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4321 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4381 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4441 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4501 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4561 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4621 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4681 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4741 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4801 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4861 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4921 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4981 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5041 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5101 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5161 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
