LOCUS       sequence-466372-        4824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3127m 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 4824)
  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 4824)
  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: AiP20083 - pAAV-AiE3127m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   146..275
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(337..925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(417..436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1096..1887,1888..1956))
                     /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     1719..1738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1957..2061)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2129..2147
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2185..2207)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2307..2326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2343..2798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2480..2501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2692..2711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2873..3013
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2884..3013
                     /label=AAV2 ITR
     polyA_signal    3131..3242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3429..3449
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3542..4252,4253..4255,4256..4258))
                     /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(3584..3605)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3933..3952
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4192..4213
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4255..4264
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tatagactct actggtgaga gagtagcttt gtcatctgct gaaccggagc tctgaaaaag
       61 agttccagct tgcattttca ggttattcct atagactcag gcttctaatg caaagcagag
      121 agcagactgc aacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct
      181 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc
      241 ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa
      301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      961 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1021 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1081 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1141 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1201 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1261 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1321 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1381 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1441 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1501 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1561 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1621 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1681 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1741 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1801 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1861 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1921 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1981 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2041 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2101 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     2161 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2221 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2281 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     2341 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2401 attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
     2461 gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2521 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2581 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2641 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2701 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2761 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     2821 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
     2881 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2941 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     3001 actaggggtt cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc
     3061 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
     3121 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
     3181 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
     3241 agtcgaagat ctctcgagcc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     3301 agcgggcaag gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca
     3361 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     3421 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     3481 ttttgtaatc cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg ccgtcgactt
     3541 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     3601 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactggtag ctcaggtagt
     3661 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     3721 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt
     3781 tcttctgctt gtcggcggtg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     3841 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     3901 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     3961 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     4021 tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta gcccagggtg gtcacgaggg
     4081 tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc agcttgccgt
     4141 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     4201 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     4261 tggcgaccgg tagcgctagc ttcgaaagat ctggatccca gaagcaaatg taagcaatag
     4321 atggctctgc cctgactttt atgcccagcc cgagctccgc cggcgggtct gcctctggct
     4381 tgaatcttga acttaccagt ttggctagtg atccccagag tccctgcctc tgactcttct
     4441 gcaccgaaac tgcacaccca gcttttgggg ggttcagggg aatctgatat caggttttca
     4501 atcttgtatg acaagcactt tactgaccaa gccacgtgct tagccccttt ccagggtgtg
     4561 ggacaatcac tcctgcaaga tcccatcccc tcatctgcac agagcttacc aagctgcagg
     4621 aacctctgag caagcttcct ggcatattgg aacaagccaa gtatttcaaa atagctactt
     4681 tagcagttcc tcaaagttat aggaaagcaa agttcaccag gatagaaact ccaagtcaga
     4741 ggattcagag ttgacttttc tacttatttg tatgaatttc attcgtggtt ttcagtactt
     4801 tagaaaccaa aaaactccag aacg
//
