LOCUS       sequence-432538-        5642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2119m 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 5642)
  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 5642)
  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: AiP1421 - pAAV-AiE2119m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(135..156)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     347..366
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   528..668
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   539..668
                     /label=AAV2 ITR
     polyA_signal    786..897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1084..1104
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1197..1907,1908..1910,1911..1913))
                     /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(1239..1260)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1588..1607
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1847..1868
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1910..1919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3443..3572
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3634..4222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3714..3733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4393..5184,5185..5253))
                     /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     5016..5035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5254..5358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5426..5444
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5482..5504)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5604..5623
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5640..5642,1..453))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
       61 ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
      121 agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
      181 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
      241 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
      301 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
      361 aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
      421 ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg
      481 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct
      541 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
      601 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
      661 gggttcctgc ggccgctcgg tccgcacgtg agatctcatg cctgctattg tcttcccaat
      721 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
      781 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
      841 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
      901 aagatctctc gagccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
      961 gcaaggcagg cggcgatgag ttccgccgtg gcaagaacta accaggattt atacaaggag
     1021 gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat
     1081 ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg
     1141 taatccagag gttgattatg atatcgaatt cgcggccgcg gcgcgccgtc gacttacttg
     1201 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     1261 tgatcgcgct tctcgttggg gtctttgctc agcttggact ggtagctcag gtagtggttg
     1321 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1381 tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc
     1441 tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     1501 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     1561 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1621 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     1681 tggtcggggt agcgggcgaa gcactgcacg ccgtagccca gggtggtcac gagggtgggc
     1741 cagggcacgg gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg
     1801 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     1861 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     1921 accggtagcg ctagcttcga aagatctgga tcccagaagc aaatgtaagc aatagatggc
     1981 tctgccctga cttttatgcc cagcccgagc tccgccggcg cagcattgtt gcatcaagtc
     2041 agagcagact ttctgagggc tgagttccca aggttccact gcacccactt gctcctgact
     2101 cggggcttca cctcgctgtg tctgccccag ggaggcgctg ggccagcagc ccagttgctc
     2161 aatatgtgca ctctctattc taactctccc cggaaactgc accatgcacg gcgctggcgc
     2221 cctccagcac ccgtgttaga aagcacaaat aataaattat caggggaaca aagatagagc
     2281 tcatgaagaa acgatcgctc tgtgggtaaa tagaacccca ctgtgtatta aacaaaaagt
     2341 aaaccccatt ttcttttgga tagttaagtt gccatcaatt caaacagaga acaaggccct
     2401 ggacgtagtt aatagtccgc agactggcca accccggctt tgaaaacgtt tcagctctcc
     2461 taaagtgctg cttgtaccca attataactc actactcacc atcttcataa tttaaatctg
     2521 tctgcccctc aggggggaaa attttttgct aatcaaagcc atgaatgcac ctctttcctt
     2581 ttgacagcat tcaattaaag ctaaatggtc ggttttgaat gagttactgt gaagaataaa
     2641 ggcattaatg aggtaatttc attatggaga ctcgtccagg cctatgtgaa aaacgaggga
     2701 actaaatgga gggcaacgct gctgagggaa ttagggtttc ttccacccta attaatgttt
     2761 gggggcattt agtacaggca ccaaattaac tgctccaaag aactctcatt gaaaaaaaaa
     2821 atacattgtt actctctcct gtaattagaa gacagtacaa ggagaaccct ctgggagtat
     2881 aaagctggcc tttaacttgt tctgagggcc accaaaaaca cacgattagg gatctggaat
     2941 ttaaggaagc aggcgtacaa tatgggattg tttaggtcct tggactctcc cagaagtaaa
     3001 caggtagcct gtgaatgagg aattatgaga ctcctggggg gcctgtcttc ccagccaaac
     3061 agtcttaaat gcatccaaaa ggaagaaaga aaggtgaata ggcccagtgt gggtcgctgg
     3121 gagaattctc aaagcctgtc ttaagtgatt acatgatgtt gctcccgccc ctctcactga
     3181 gttatcttta cacggccttt gtctgctcta attggagctg atgaggtgct aattggaggc
     3241 ctttttaaaa agcccacgac tttaatcatt agaaaagcgg gggaatctga cggagtggag
     3301 ttaatgagac cggagagaag agggaaccga atgtgtagct caacaaaaat aaaactggtc
     3361 ttgcatgtgt gcagggggga ggtttgatgg aaacgggaaa ggagtcgccg gcgattatta
     3421 ctcttacaac gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3481 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc
     3541 agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc
     3601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4681 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4981 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5401 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5461 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5521 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5581 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
     5641 aa
//
