LOCUS       sequence-465216-        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2554m 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 4888)
  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 4888)
  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: AiP1926 - pAAV-AiE2554m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4888
                     /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      900..909
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(906..908,909..911,912..1622)
                     /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(951..972)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1212..1231)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1559..1580
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1715..1735)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1922..2033
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2151..2291
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2151..2280
                     /label=AAV2 ITR
     rep_origin      2366..2821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2453..2472)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2663..2684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2838..2857)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2957..2979
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3017..3035)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3103..3207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3208..3276,3277..4068)
                     /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(3426..3445)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4239..4827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4728..4747
                     /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 gcggccgcat tgttttgttt ttctctctgg gatgagagga gaatgaagaa
      181 tgcaccttat tagaatcaga gaacaagaag gagttgagcc ctctgactcc tcttagaatt
      241 ttaaagtaag ccatataggg gaggaaaaaa aatacttctc aaagttgaca caaatgttta
      301 ggggaaatca cagaatattg tttaaaggtt ttagctgaaa gcccgatatt tgagaattag
      361 tgaagaggtt taatattgtt gtgtagtttg tgtagcttgc tgtttgtggg aacttgcaga
      421 tacaatgcat acagactcgt cggtgcccct ccgcaatcac acacactggc tgctggatat
      481 ggcagtgata tcacagagag tgagttagca tctgttcagc cctgagtcag tgacatatgc
      541 tgtgtcttat tttgactagc tagtagctcg agatttaaga aaaatatgat cttagaaaaa
      601 tatgacaaaa ttgcatttta gtactccgag atttgtgttt gtggggctat tctaaacaca
      661 aatcctttca tttgaaatag tgctgaaatt atgtcatctt gcttttcctg taaatttttg
      721 gaagagactc atctcttcct tttttattgt ggagtaccag tattatttac attgaacttc
      781 ctacctcaaa tgtgcctgta cttacggagc tcgggctggg cataaaagtc agggcagagc
      841 catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg
      901 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      961 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1021 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     1081 ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca
     1141 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1201 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1261 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1321 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     1381 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     1441 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1501 accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     1561 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1621 agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca
     1681 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     1741 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     1801 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     1861 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg
     1921 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     1981 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2041 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2101 tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct
     2161 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2221 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2281 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2341 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2401 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     2461 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2521 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2581 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2641 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     2701 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     2761 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     2821 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     2881 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2941 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3001 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3061 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3121 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3181 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3241 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3301 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3361 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3421 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3481 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3541 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3601 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3661 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3721 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3781 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3841 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3901 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3961 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4021 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4081 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4141 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4201 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4261 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4321 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4381 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4441 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4501 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4561 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4621 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4681 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4741 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4801 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4861 gccttttgct ggccttttgc tcacatgt
//
