LOCUS       sequence-471248-        4818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2560m 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 4818)
  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 4818)
  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: AiP1951 - pAAV-AiE2560m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4818
                     /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      830..839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(836..838,839..841,842..1552)
                     /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(881..902)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1142..1161)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1489..1510
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1645..1665)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1852..1963
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2081..2221
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2081..2210
                     /label=AAV2 ITR
     rep_origin      2296..2751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2383..2402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2593..2614
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2768..2787)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2887..2909
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2947..2965)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3033..3137
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3138..3206,3207..3998)
                     /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(3356..3375)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4169..4757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4658..4677
                     /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 gcggccgcag agcaaacaac aacaataaca gtaaagagcc cagcctcagg
      181 gattttgtta cagctacaca gaatgtacaa agcctctctg aatttcattt cactcaaaca
      241 ccttttcttt taaaagcttt ttggtgctgg gcaaccactg aactacttcc caggcaaatt
      301 ctgtcttaat ctttgcgttg cgcgtctccc ggagaaaccc agtgtttcgg gaccatgcag
      361 acctggcttc tccatgtttg acagctattg gttagtttat ctctttcatg tcccatttca
      421 ctacaaacac cagaaataaa ccagatggca tcttcaacat tctgcctgga aaccttgtaa
      481 ctacatcacc cggctcatta ggcgcctttc tggctgttca tgttactgtg agtgggtgtt
      541 gcaaactctg ttactgctgc aaggatcctg tcttcgccat ttctagtgag cgtgtcctct
      601 tacacccttg ctgagggctc tttgacattt ttccttagga ctcttggtcc tttatcttcc
      661 ttctcagttt ttcgtccctt ggctatgtgg tcccaaggcc atgctcaaac tttgggtttt
      721 cattactgta gctgcggagc tcgggctggg cataaaagtc agggcagagc catctattgc
      781 ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt
      841 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      901 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      961 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1021 gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1081 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1141 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1201 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1261 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1321 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     1381 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1441 gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1501 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga
     1561 cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga
     1621 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1681 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     1741 atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     1801 aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt
     1861 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1921 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1981 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2041 atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     2101 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2161 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2221 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     2281 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2341 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     2401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     2521 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2581 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     2641 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     2701 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     2761 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2821 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2881 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2941 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3001 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3061 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3121 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3181 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3241 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3301 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3361 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3421 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3481 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3541 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3601 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3661 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3721 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3781 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3841 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3901 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3961 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4021 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4081 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4141 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4741 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4801 ggccttttgc tcacatgt
//
