LOCUS       sequence-473489-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0195h 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 5418)
  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 5418)
  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: AiP11688 -
            pAAV-hsA2-AiE0195h-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5418
                     /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      1455..1464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1461..1463,1464..1466,1467..2177)
                     /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(1506..1527)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1767..1786)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2114..2135
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2245..2265)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2452..2563
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2681..2821
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2681..2810
                     /label=AAV2 ITR
     rep_origin      2896..3351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2983..3002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3193..3214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3368..3387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3487..3509
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3547..3565)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3633..3737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3738..3806,3807..4598)
                     /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(3956..3975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4769..5357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5258..5277
                     /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 tttgtttaga
      361 gaagagcagg aagaaacact tttccttctc actcaaggtt ggactgtaag taattttaac
      421 aactcctgtc tgccccacgc ctttagacaa gagcaatttg caactctagg gctgcttaca
      481 ttgcaaagat ctctattctc tcccatccca gagacaaact ctgctgtcaa tatgactttg
      541 ctgagagaaa ttagttcatg tgtctattaa aagattgcat acaaatgttc aaagcatcct
      601 tatttctaat agccccaaaa tggaaactag cccagtgtgc atcattggga gaatggataa
      661 aaacttgtgg gatgtgatgc aacggaaggc tcttcagcaa taaaaggaaa aggaaatcgt
      721 gattgtgaca tgtgccatgg ctgaatttca tagcaacaag gatgagagaa atgctaaaca
      781 taagagaatg ctgctccaag tttgaatctg aacttcttgc tttgtgtacc aagccattct
      841 tgcccacatc caatctccaa tatcgttctc catttcccaa gccttagtct tttccaagtg
      901 gaatgacatg ctcaaatcag ttttctaaat atggttaata cactcatatt tgtgactatg
      961 gctattcatt catttagaaa tatttgaatg tctgtgctag ccatcacact gatacaatgg
     1021 tgacatcata gtgaagaaga caacgcagtc ttatggaact ctgggaggga tgaaggtaga
     1081 agctatgggt catgcatctt atgctttgct ggatctttag tccctggatc gttgtagaca
     1141 attgatagac ttgttaaatt gacacatgag tgcattgata aacaaattca tttgaacaga
     1201 gcaaaagatt gagattgatt cagttattca gttgaaaaca gttgagaggc tgtctgtgtg
     1261 gcgaattcga tatcataatc aaccataggt accgagctcg ggattcagcc gggagcttag
     1321 ggaggggagg tcacttcata agggcctggg gggggagttg gagccacgag tcgtccagcc
     1381 ggagccccgt gtggctgagc tccggcctca gaagcatccc cgggttggat ccttcgaagc
     1441 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1501 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1561 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     1621 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
     1681 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1741 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1801 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1861 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1921 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     1981 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2041 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     2101 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2161 tggacgagct gtacaagtaa gtcgacatca taatcaacct ctggattaca aaatttgtga
     2221 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2281 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2341 atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2401 aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt
     2461 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2521 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2581 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2641 atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     2701 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2761 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2821 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     2881 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2941 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3001 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3061 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3121 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3181 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     3241 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     3301 tttaacaaaa atttaacggg aattttaaca aaatattaac gtttacaatt ttatggtgca
     3361 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3421 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3481 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3541 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3601 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3661 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5401 ggccttttgc tcacatgt
//
