LOCUS       sequence-461979-        4758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2272m 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 4758)
  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 4758)
  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: AiP1605 - pAAV-AiE2272m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /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      770..779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(776..778,779..781,782..1492)
                     /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(821..842)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1082..1101)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1429..1450
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1585..1605)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1792..1903
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2021..2161
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2021..2150
                     /label=AAV2 ITR
     rep_origin      2236..2691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2323..2342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2533..2554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2708..2727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2827..2849
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2887..2905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2973..3077
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3078..3146,3147..3938)
                     /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(3296..3315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4109..4697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4598..4617
                     /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 gcgttgtaag agtaataatc gccggcgcca tccatctcaa
      181 ggccacaggc tgagcagata ttttccatct ccacagctga atgcagacat tccagagagc
      241 actgcttcct ctctcctgag cttctaggca ggcgagggtc tcaagcacaa catctgtgcc
      301 taggccccag ctctctgtca cagggaagga agagacaaag cccagggtct tgcagccagg
      361 cctgagaggc ccagtcaggg ggagcaagcc aggcctccca gccagcacca ttcacaccag
      421 gcccttccca cttcctcttt cccctttctc cccatgacaa catcttgaac cagtgccttc
      481 actggcaagc ccaagcactc tggggttttg tagttctgag gaggctccca cgctcagctg
      541 tgcacacagc ccttggacaa gctgctgagg ttgcctgtgg tactcggggc aaaacacgaa
      601 tctccagccc acacatgttc tctgaggcct ctgcttcagg gacggaggtg aggtgtccct
      661 ggccctcccg ccggcggagc tcgggctggg cataaaagtc agggcagagc catctattgc
      721 ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt
      781 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      841 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      901 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      961 gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1021 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1081 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1141 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1201 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1261 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     1321 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1381 gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1441 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga
     1501 cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga
     1561 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1621 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     1681 atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     1741 aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt
     1801 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1861 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1921 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1981 atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     2041 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2101 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2161 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     2221 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2281 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     2341 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2401 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     2461 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2521 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     2581 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     2641 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     2701 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2761 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2821 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2881 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2941 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3001 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3061 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3121 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3181 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3241 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3301 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3361 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3421 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3481 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3541 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3601 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3661 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3721 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3781 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3841 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3901 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3961 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4021 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4501 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4681 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4741 ggccttttgc tcacatgt
//
