LOCUS       sequence-470886-        5514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0023h_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 5514)
  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 5514)
  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: AiP11521 -
            pAAV-hsA2-AiE0023h_3xC2-minRho-SYFP...
FEATURES             Location/Qualifiers
     source          1..5514
                     /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      1551..1560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1557..1559,1560..1562,1563..2273)
                     /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(1602..1623)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1863..1882)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2210..2231
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2341..2361)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2548..2659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2777..2917
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2777..2906
                     /label=AAV2 ITR
     rep_origin      2992..3447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3079..3098)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3289..3310
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3464..3483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3583..3605
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3643..3661)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3729..3833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3834..3902,3903..4694)
                     /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(4052..4071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4865..5453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5354..5373
                     /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 tttgttttgg
      361 tcctggttct gactcaagct gcaaccccca gaggcagaag gagagggatg catggagggg
      421 aatgtctttt tccaccaagc agctgctccc tttcacaaga ggccagttat gtaaggaagg
      481 ctgcttcctc tccccttgct aaatcttttc tagtgcagca taaggccaga gctagggaaa
      541 gtccacacgg gaataaaaag agcagattgc taagcacaga agccatttat aactaaaacg
      601 tctcagctca gcttagggac gcgtcctagg tcacgactcc acccctccag gaacccctag
      661 tgagggagtg gtgggagcag agccctcatc acacagactt ggtcctggtt ctgactcaag
      721 ctgcaacccc cagaggcaga aggagaggga tgcatggagg ggaatgtctt tttccaccaa
      781 gcagctgctc cctttcacaa gaggccagtt atgtaaggaa ggctgcttcc tctccccttg
      841 ctaaatcttt tctagtgcag cataaggcca gagctaggga aagtccacac gggaataaaa
      901 agagcagatt gctaagcaca gaagccattt ataactaaaa cgtctcagct cagcttaggg
      961 acgcgtccta ggtcacgact ccacccctcc aggaacccct agtgagggag tggtggtggg
     1021 tgagtaagag taggtggcaa ctggtcctgg ttctgactca agctgcaacc cccagaggca
     1081 gaaggagagg gatgcatgga ggggaatgtc tttttccacc aagcagctgc tccctttcac
     1141 aagaggccag ttatgtaagg aaggctgctt cctctcccct tgctaaatct tttctagtgc
     1201 agcataaggc cagagctagg gaaagtccac acgggaataa aaagagcaga ttgctaagca
     1261 cagaagccat ttataactaa aacgtctcag ctcagcttag ggacgcgtcc taggtcacga
     1321 ctccacccct ccaggaaccc ctagtgaggg agtggtggga attcgatatc ataatcaacc
     1381 ataggtaccg agctcgggat tcagccggga gcttagggag gggaggtcac ttcataaggg
     1441 cctggggggg gagttggagc cacgagtcgt ccagccggag ccccgtgtgg ctgagctccg
     1501 gcctcagaag catccccggg ttggatcctt cgaagctagc gctaccggtc gccaccatgg
     1561 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1621 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1681 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1741 tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1801 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1861 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1921 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1981 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     2041 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     2101 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2161 tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2221 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg
     2281 acatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2341 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2401 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     2461 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2521 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2581 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2641 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2701 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2761 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2821 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2881 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2941 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3001 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3061 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3121 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3181 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3241 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3301 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3361 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgggaatt
     3421 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3481 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3541 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3601 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3661 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3721 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3841 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3901 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3961 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4021 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4081 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4141 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4201 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4261 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4321 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4381 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4441 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4501 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4561 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4621 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4681 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4741 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4801 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4861 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4921 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4981 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5041 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5101 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5161 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5221 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5281 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5341 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5401 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5461 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
