LOCUS       sequence-470792-        4742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0357h_3xC2 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 4742)
  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 4742)
  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: AiP15301 -
            pAAV-AiE0357h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4742
                     /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      754..763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(760..762,763..765,766..1476)
                     /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(805..826)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1066..1085)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1413..1434
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1569..1589)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1776..1887
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2005..2145
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2005..2134
                     /label=AAV2 ITR
     rep_origin      2220..2675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2307..2326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2517..2538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2692..2711)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2811..2833
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2871..2889)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2957..3061
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3062..3130,3131..3922)
                     /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(3280..3299)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4093..4681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4582..4601
                     /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 gcgtattgag acttgggttt atctgcctca catcaggtac
      181 atcaagttct ggatggctgc ccactggcca gagacatgag gtggacggct cccttttgct
      241 gccctgggaa ggccttctgc tggcttcggc cccactgagc aaagtctgct tgttcactgg
      301 agttcacaca gactccattg agacttgggt ttatctgcct cacatcaggt acatcaagtt
      361 ctggatggct gcccactggc cagagacatg aggtggacgg ctcccttttg ctgccctggg
      421 aaggccttct gctggcttcg gccccactga gcaaagtctg cttgttcact ggagttcaca
      481 cagactccat tgagacttgg gtttatctgc ctcacatcag gtacatcaag ttctggatgg
      541 ctgcccactg gccagagaca tgaggtggac ggctcccttt tgctgccctg ggaaggcctt
      601 ctgctggctt cggccccact gagcaaagtc tgcttgttca ctggagttca cacagactcc
      661 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      721 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      781 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      841 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
      901 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
      961 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1021 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1081 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1141 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1201 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1261 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1321 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1381 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1441 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1501 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1561 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1621 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     1681 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1741 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     1801 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1861 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1921 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     1981 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2041 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2101 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     2161 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     2221 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2281 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2341 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2401 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2461 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2521 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2581 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2641 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     2701 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2761 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2821 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2881 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2941 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3001 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3061 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3121 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3181 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3241 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3301 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3361 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3421 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3481 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3541 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3601 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3661 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3721 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3781 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3841 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3901 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3961 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4021 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4081 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4141 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4201 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4261 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4321 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4381 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4441 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4501 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4561 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4621 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4681 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4741 gt
//
