LOCUS       sequence-461974-        4843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2578m 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 4843)
  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 4843)
  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: AiP1891 - pAAV-AiE2578m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     RBS             384..392
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      855..864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(861..863,864..866,867..1577)
                     /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(906..927)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1167..1186)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1514..1535
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1670..1690)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1877..1988
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2106..2246
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2106..2235
                     /label=AAV2 ITR
     rep_origin      2321..2776
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2408..2427)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2618..2639
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2793..2812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2912..2934
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2972..2990)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3058..3162
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3163..3231,3232..4023)
                     /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(3381..3400)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4194..4782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4683..4702
                     /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 agagagagag aaagagagag agggagggag agagaggaga
      181 attcagtcaa agctgagttc aatatggttg ggttctaatc tcgcatttct tagcagaggc
      241 acatttttct tttggatctg aaagcactgc tggctgctat ctgcggtaag aaataccgtc
      301 ctgaggcaga tgtctatttc ctctgccagg ctggtgtgga agccgaccct gtttcctctt
      361 ccatctggat gctggggaag cagtaaggag gtgtcaagca ttctgcccca aacattcttt
      421 ctcatccgcg tgctctcgtg ctagctaaca tcttttaaaa tagcagcaaa tggctccgaa
      481 agagataact taagcctgtg aagccaggac aacagcgagg gaagcggaat ctctgaattt
      541 gtgatccatg cccattctga catctaggca agctttgaac tacaagtcct ggcttcgaat
      601 ctcagctctt aagcagcttg ccgtggacag tattggaggg tgtaatctct gaattgcatt
      661 gcagattcat taatatgaat gtgaaacacc cacagtctga aggagaggga gagatgtttc
      721 tgcccagttt ctattgggaa ggagtcacct agttttggcc ggagctcggg ctgggcataa
      781 aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc
      841 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      901 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      961 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     1021 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
     1081 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1141 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1201 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1261 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1321 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     1381 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1441 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     1501 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1561 tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc
     1621 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1681 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1741 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     1801 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     1861 ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1921 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1981 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2041 agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg
     2101 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     2161 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     2221 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     2281 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     2341 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     2401 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2461 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2521 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2581 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2641 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     2701 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2761 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2821 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2881 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2941 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3001 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3061 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3121 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3181 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3241 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3301 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3361 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3421 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3481 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3541 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3601 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3661 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3721 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3781 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3841 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3901 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3961 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4021 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4081 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4141 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4201 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4261 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4321 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4381 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4441 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4501 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4561 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4621 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4681 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4741 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4801 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
