LOCUS       sequence-441077-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2103m 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 5807)
  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 5807)
  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: AiP1287 - pAAV-AiE2103m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..94)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(754..1545,1546..1614))
                     /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     1377..1396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1615..1719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1787..1805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1843..1865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1965..1984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2001..2456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2138..2159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2350..2369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2531..2671
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2542..2671
                     /label=AAV2 ITR
     polyA_signal    2789..2900
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3087..3107
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3200..3910,3911..3913,3914..3916))
                     /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(3242..3263)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3591..3610
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3850..3871
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3913..3922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5611..5740
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(5802..5807,1..583))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
       61 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      421 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1741 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     1801 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     1861 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     1921 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     1981 tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     2041 tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa tcccttataa
     2101 atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca agagtccact
     2161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2221 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt
     2401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta
     2461 tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2521 atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
     2581 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     2641 ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac gtgagatctc
     2701 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
     2761 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
     2821 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
     2881 ctggcaacta gaaggcacag tcgaagatct ctcgagccac ggaattgtca gtgcccaaca
     2941 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa
     3001 ctaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     3061 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     3121 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atgatatcga attcgcggcc
     3181 gcggcgcgcc gtcgacttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     3241 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg
     3301 actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     3361 tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga
     3421 agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt
     3481 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     3541 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     3601 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     3661 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc
     3721 ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct
     3781 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     3841 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     3901 cgcccttgct caccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga
     3961 agcaaatgta agcaatagat ggctctgccc tgacttttat gcccagcccg agctcgtctt
     4021 tgatttctgg gcagaacatt ttaacttaag tatcccacta tttctttttt tgcaagcgag
     4081 aagtagataa ttagatatga attcatctat taacacttat gctttctatt ttcattttca
     4141 ccaaagtaat tttctccatc agttataatg ctgaagaaag ataatttttt gtagcaaaga
     4201 aaactctttg ttatctaatc ttttgtgtaa tgaaagggaa acacaaaatg aactttgaaa
     4261 aaggggactt ttctgttggt aattgttctg ctatgtgctg ccagttctgc attcaggcct
     4321 aatatgaagc caagaagttg tatatggata tgtttaatcc agtttttgta ttaatcacaa
     4381 taaagcatga aagacatcat tatatttcag aagaggctct gaagttaatg tagaataaaa
     4441 aaacaattct attatgtagg gtataccaac taatgatttt attttactaa aacgtgagtt
     4501 tcagatctag ccagaacatt aaaaggtact gttgagtttt caggttaacc ccatggcctg
     4561 tagaatgttt aggtttgaga tattaatttg atttaaaagt tattgctgaa gtaatttgcc
     4621 taaagatcta gttggaagag ttcacttgtg aaaactcttg tttgcacaaa tctggatgat
     4681 tgcctggata agacaggaag tggcggtggc agtggaaggg agtagccaca aggagagaag
     4741 agtaaaagtt tttctcaatt aaaattccat ggataaatac caaactgaaa ggtgtacagt
     4801 gtcctggaag tgtttctgag accctatatc acatgtagaa atgtacacac tcagcccaga
     4861 aacaatgtgc aatgaacagg aattttaagt cattctttta agtatttcag aaaaagcaat
     4921 aaccaacaat caaaatccct aaagtcctgc ccacccctgc aacttatttt acatttttgt
     4981 gatgtttgct ggtatgtctt cactctacta gcatctcaaa gaaacacttg gttcctccat
     5041 cgctcatggt gctgatgttt gtgttcagct ctttagaatt gtgttttctg gctcttgtag
     5101 agtcttgcca cactattgta gagctttgga cattgtctag tgattgctag ggaatacatg
     5161 tgtataatag tgttctatat tcatcttctc attcttttcc tgagtttctg tctgctggtt
     5221 gttattctac tttactttgt cttacttttt ataccaaatt aactcaattg attcaaaact
     5281 gaacgttggg ctagcataga tcattctctt gaaaatataa aacagaatgc acttcagcag
     5341 gagaaggagg ctggctggtc atactgtgtc caaagacagg aaatggagag tgaactgaaa
     5401 aatgggggtc agagcacaaa acatcaaaat tttccatttg tgacccactc tattaagcaa
     5461 ggctccttct tctaaaggtc taaaagtctt ctaaaacagc atcactaact ggagactaag
     5521 ttatcactaa ctacaacctg ttgagacatt tcctattcag actgaagctt tgagtgacac
     5581 tgcagatatc ctttgcacgc gtgcggccgc aggaacccct agtgatggag ttggccactc
     5641 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5701 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag
     5761 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttcca
//
