LOCUS       sequence-461964-        4696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3052m 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 4696)
  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 4696)
  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: AiP20017 - pAAV-AiE3052m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4696
                     /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      708..717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(714..716,717..719,720..1430)
                     /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(759..780)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1020..1039)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1367..1388
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1523..1543)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1730..1841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1959..2099
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1959..2088
                     /label=AAV2 ITR
     rep_origin      2174..2629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2261..2280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2471..2492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2646..2665)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2765..2787
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2825..2843)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2911..3015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3016..3084,3085..3876)
                     /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(3234..3253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4047..4635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4536..4555
                     /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 gcgtgttgtc tatatggcac tgttagcata aaaatgggaa
      181 gatttgtgtg ctaagatcac atttgtgatt ccctggctgc tgctgtgaca ttgtagtctg
      241 gtcacatttt taccccaatt ctgagcagga atgatccttt aggttgaggc ctcagacctg
      301 aacatcagca gagctgagaa agttgtatca ttcatttcta tagctctctg attgtatctt
      361 ctcctgacca actcttgtgc ttccagatgc ccagaggaca tgtgacatgg ataaacccca
      421 ggactccttc taggaagcat ttctctctga cagtttgaga cacatcatct gttattttta
      481 agtctaagct ggaagatgat ttattgtggc ttcaaagcat atgctacttt gtgttttata
      541 aatggaaact gggtgagatg tatcctctga aatgcttggt tgggctatgt ttaatcaagc
      601 cctagtcgcc ggcggagctc gggctgggca taaaagtcag ggcagagcca tctattgctt
      661 acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtga
      721 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      781 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      841 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      901 ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
      961 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1021 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1081 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1141 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     1201 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1261 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1321 gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1381 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagtcgacg
     1441 gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa atttgtgaaa
     1501 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1561 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1621 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     1681 gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct
     1741 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1801 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1861 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1921 agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     1981 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     2041 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     2101 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     2161 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2221 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     2281 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2341 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     2401 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2461 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     2521 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     2581 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     2641 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2701 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2761 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2821 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2881 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2941 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3001 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3061 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3121 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3181 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3241 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3301 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3361 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3421 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3481 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3541 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3601 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3661 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3721 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3781 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3841 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3901 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3961 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4021 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4081 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4141 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4201 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4261 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4321 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4381 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4441 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4501 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4561 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4621 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4681 ccttttgctc acatgt
//
