LOCUS       sequence-438972-        4750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2344m 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 4750)
  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 4750)
  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: AiP1762 - pAAV-AiE2344m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(127..231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     299..317
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(355..377)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     477..496
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(513..968)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(650..671)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     862..881
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1043..1183
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1054..1183
                     /label=AAV2 ITR
     polyA_signal    1301..1412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1599..1619
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1712..2422,2423..2425,2426..2428))
                     /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(1754..1775)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2103..2122
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2362..2383
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2425..2434
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3066..3195
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3257..3845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3337..3356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4016..4750,1..57,58..126))
                     /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     4639..4658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
       61 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
      121 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
      181 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
      241 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
      301 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
      361 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
      421 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
      481 atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa
      541 attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa
      601 aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa
      661 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
      721 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
      781 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
      841 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc
      901 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
      961 gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg
     1021 agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     1081 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     1141 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc
     1201 acgtgagatc tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
     1261 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
     1321 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
     1381 aacaacagat ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt
     1441 cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg
     1501 ccgtggcaag aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     1561 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     1621 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc
     1681 gaattcgcgg ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga
     1741 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     1801 tgctcagctt ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     1861 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt
     1921 ggcggatctt gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt
     1981 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     2041 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     2101 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     2161 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact
     2221 gcacgccgta gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     2281 tgcagatcag cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     2341 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     2401 tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat
     2461 ctggatccca gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc
     2521 cgagctccgg tgtagcagtg tgttaagtag gtagtaggca gcaggcaaat gcctccacag
     2581 gcctccactc cacgggccac cactcactca ctcacttagc tatgttctaa gacctaaatc
     2641 gtgtgctatt gtcacccaca gcacactcag gaaaatctag actccaaaaa gttaaagcct
     2701 tagctaaggt tttatggaaa gtaaacagca gagtcagtat tttcttcctt ccccacagct
     2761 ttctaagaaa aggatgttgt gcagtttcca ttgaatacat cacaagctgt tagcaaaaat
     2821 gtttcttgct ggaatataag caaaaggtct tgggaggcat ggaggaagga cagacagcat
     2881 attcttcaag cattactaat tgtgtgaaca ctctaaaaca cagaaaaccc ctcccagtcc
     2941 cagagactgg catatcccat ttctgtcttt taccactagc tgttttcata aacgctcatc
     3001 attttggtgt tctaaacacc ctcagagtac agaccgatta ttactcttac aacgcgtgcg
     3061 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3121 tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc
     3181 gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3241 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4741 cgatgtaacc
//
