LOCUS       sequence-491908-        4678 bp    DNA     circular SYN 05-NOV-2025
DEFINITION  AiE0086h 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 4678)
  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 4678)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 5, 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: AiP12674 - pAAV-AiE0086h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4678
                     /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      690..699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(696..698,699..701,702..1412)
                     /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(741..762)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1002..1021)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1349..1370
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1505..1525)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1712..1823
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1941..2081
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1941..2070
                     /label=AAV2 ITR
     rep_origin      2156..2611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2243..2262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2453..2474
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2628..2647)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2747..2769
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2807..2825)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2893..2997
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2998..3066,3067..3858)
                     /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(3216..3235)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4029..4617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4518..4537
                     /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 gcggccgcac gcgtacagcc ttttcctcct ctgacattcc tgacctcccc
      181 gtggcctgcc ttcccctgtc tccttggccg agccatcctc tgtcaccttc actgtttcca
      241 ctaggttatt gaggcctttt ggtcatgagc tcctgccacc aaatcagtgc ctgaggctgc
      301 tgcataacaa tctccttcct tgattagagc atcttaatta tgctccagga atttgcttaa
      361 acagattaat cattctggag gacagattta gaagctgtgc tagctgaggg atctgctctg
      421 agtgtccccg gggagattct ggtgacaacc gcctgttggg cctttgccaa ggccttgggt
      481 gcctcggaga ctcggacact ggctggaagc tctacgcacc cccgaaggct tctctttgct
      541 cctctgggcc tgtagagcag gctctgaaac tcttggatgt gaaacgtggc aggagtgagc
      601 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
      661 cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
      721 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      781 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
      841 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
      901 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      961 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1021 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1081 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1141 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1201 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1261 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     1321 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1381 ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa
     1441 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1501 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1561 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     1621 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     1681 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     1741 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1801 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1861 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     1921 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     1981 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2041 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2101 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2161 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2221 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2281 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2341 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     2401 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2461 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     2521 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2581 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     2641 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2701 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2761 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2821 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2881 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2941 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3001 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3061 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3121 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3181 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3241 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3301 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3361 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3421 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3481 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3541 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3601 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3661 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3721 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3781 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3841 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3901 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3961 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4021 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4081 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4141 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4201 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4261 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4321 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4381 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4441 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4501 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4561 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4621 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
