LOCUS       sequence-432575-        4660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2557m 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 4660)
  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 4660)
  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: AiP1865 - pAAV-AiE2557m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     86..107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(261..280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     380..402
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(440..458)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        526..630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(631..699,700..1491)
                     /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(849..868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1662..2250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2151..2170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2312..2441
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2983..2992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2989..2991,2992..2994,2995..3705)
                     /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(3034..3055)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3295..3314)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3642..3663
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3798..3818)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4005..4116
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4234..4374
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4234..4363
                     /label=AAV2 ITR
     rep_origin      join(4449..4660,1..244)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4536..4555)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
       61 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
      121 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
      181 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
      241 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
      301 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      361 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      421 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      481 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      541 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      601 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      661 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      721 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      781 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      841 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
      901 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      961 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1021 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1081 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1141 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1201 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1261 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1321 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1381 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1441 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1501 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1561 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1621 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1681 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1741 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1801 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1861 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1921 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1981 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2041 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2101 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2161 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2221 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2281 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
     2341 actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     2401 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca caagaaagaa
     2461 tttggctttt gaatgttatt ccaagtcctc tgcctctttg aatgtattcg tcttacttcc
     2521 ttatgctaag atattctccc aactgataaa tttcactgct gtgggcaagt ttttcctcct
     2581 tcagttttag cgtgtgatat ctccctatgt ctcagcataa gtctggataa gttccatctc
     2641 actgcttcca caaactattt cagagctcct ttctcatggc actcatccca gtgaattcat
     2701 ttttttcaga tttattgaag tatagttgac atacaatcaa cagcacattg tgatgtgtat
     2761 gatttaatga gctctgacac atgtgtacag ccataaatca tcatcacaaa ttaagataat
     2821 gaccttttcc caccaaccaa agcagtttct tcccatcctt tcttacttcc tcttgtctct
     2881 cctcatccgg agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     2941 tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag
     3001 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3061 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3121 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3181 ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     3241 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3301 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3361 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3421 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     3481 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     3541 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     3601 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3661 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg
     3721 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
     3781 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3841 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3901 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3961 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     4021 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4081 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4141 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4201 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     4261 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4321 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4381 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4441 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4501 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4561 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4621 ccgatttagt gctttacggc acctcgaccc caaaaaactt
//
