LOCUS       sequence-466262-        4964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2151m 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 4964)
  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 4964)
  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: AiP1434 - pAAV-AiE2151m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(74..865,866..934))
                     /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     697..716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(935..1039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1107..1125
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1163..1185)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1285..1304
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1321..1776)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1458..1479)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1670..1689
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1851..1991
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1862..1991
                     /label=AAV2 ITR
     polyA_signal    2109..2220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2407..2427
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2520..3230,3231..3233,3234..3236))
                     /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(2562..2583)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2911..2930
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3170..3191
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3233..3242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4088..4217
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4279..4867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4359..4378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
       61 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
      241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
      301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
      361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
      421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
      481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
      541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
      601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
      661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
      721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
      781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
      841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
      901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
      961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1021 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1081 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1141 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1201 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     1261 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     1321 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     1381 tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     1441 tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     1501 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     1561 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     1621 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     1681 cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     1741 cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg
     1801 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca
     1861 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     1921 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     1981 taggggttcc tgcggccgct cggtccgcac gtgagatctc atgcctgcta ttgtcttccc
     2041 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     2101 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     2161 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     2221 tcgaagatct ctcgagccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag
     2281 cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa ctaaccagga tttatacaag
     2341 gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg
     2401 tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt
     2461 ttgtaatcca gaggttgatt atgatatcga attcgcggcc gcggcgcgcc gtcgacttac
     2521 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     2581 atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actggtagct caggtagtgg
     2641 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     2701 agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc
     2761 ttctgcttgt cggcggtgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     2821 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     2881 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     2941 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     3001 atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc ccagggtggt cacgagggtg
     3061 ggccagggca cgggcagctt gccggtggtg cagatcagct tcagggtcag cttgccgtag
     3121 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     3181 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg
     3241 gcgaccggta gcgctagctt cgaaagatct ggatcccaga agcaaatgta agcaatagat
     3301 ggctctgccc tgacttttat gcccagcccg agctccgccg gcgaaagagt taaggaaagt
     3361 gctgggaaaa aaaaaaaaaa ccatgctgtg tcctactacc tcagtgagtg actgtattag
     3421 gtttctgttt ctgcctgtca tccatggtgg aaggaggtcc tcagattagc attccaaatc
     3481 ctggagtctg agagggagct gggacttggg tggtgctgtt tcaagcattt gcagtttgct
     3541 cctctctggc tccttaacca agtgtgcagt ctcttttcca agaacccttg tgtttgggaa
     3601 gtgctccgca gtggtgaaaa gtttgaacag gaaagattaa tcaaaggttg ggtctttcat
     3661 gctcagcgca gaacatgttt agttcttgga tggaagctct tcgccaggaa actagttaaa
     3721 taatattcag gcgatgaatc tgagctccag ctgcaactaa actaaaggtc agcttagtct
     3781 caactgattc ctctcacatc tgtccagaaa cactgtgcca caaggatttt cttcctattt
     3841 cggtatggct cctgaagcgt gagagttccc taaacagttg gggtttttct ttgttgttta
     3901 catgagtggc tattttgaga taaggtctca cataacccag cctaggcata aagtttgtta
     3961 cgtaggtgag aatggacttg aacttctgat gaccctcttg cttttgcctc ctaagttgga
     4021 agattgcaag catgctccac catgccctgc cgccggcgat tattactctt acaacgcgtg
     4081 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4141 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     4201 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4681 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttca
//
