LOCUS       sequence-466247-        4963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2138m 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 4963)
  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 4963)
  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: AiP1367 - pAAV-AiE2138m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      376..385
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(382..384,385..387,388..1098)
                     /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(427..448)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(688..707)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1035..1056
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1191..1211)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1398..1509
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1627..1767
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1627..1756
                     /label=AAV2 ITR
     rep_origin      1842..2297
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1929..1948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2139..2160
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2314..2333)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2433..2455
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2493..2511)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2579..2683
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2684..2752,2753..3544)
                     /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(2902..2921)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3715..4303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4204..4223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4365..4494
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgttgggatt gttgtgatcc atggctttct cccaaaccaa atcatccgcc ccggagtttc
       61 cataaacctt tctcaagggg tgtgagctac cagaaagatc atggtgtgtc ctgggtgctg
      121 gcctttgaga acccacctag ccctctccag cctctccctc aggctggagg ctggaggaag
      181 attatttttg tacttgagta actgcgtcaa ccttaggagt aataacttgg ctatgactct
      241 atttatttat tcacaaagaa agtgcccttc tcctggagaa ttgagctcgg gctgggcata
      301 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
      361 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      421 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      481 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
      541 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
      601 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      661 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      721 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      781 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
      841 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      901 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      961 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     1021 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1081 atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat
     1141 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1201 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1261 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     1321 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     1381 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1441 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1501 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1561 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     1621 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     1681 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     1741 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     1801 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     1861 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     1921 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1981 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2041 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2101 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2161 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     2221 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2281 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2341 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     2401 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2461 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     2521 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2581 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2641 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2701 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2761 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2821 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2881 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2941 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3001 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3061 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3121 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3181 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3241 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3301 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3361 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3421 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3481 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3541 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3601 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3661 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3721 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3781 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3841 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3901 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3961 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4021 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4081 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4141 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4201 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4261 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4321 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     4381 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     4441 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     4501 gcacgcgtcc tcagctcaat cctggataca taaaagtggt aatgtagccc atatgtttac
     4561 agggtaggct cgataaggac gatagataca tatacatcta ctgtgttacc tggttctagg
     4621 acccacttca gtataggaac aagactaaac agaaattggg ccgcatccca ttgcacactt
     4681 ccttagctct catctcctct actccgagac acggagtgct ttccacagca accacgtagg
     4741 tgggagctgg agtcatttga ggtcttaggg ccaaggcggt gcttcgcctc aaggctatgc
     4801 agtttccaag tccagacatc tggattttag ggtctataat ggaaactcag tcggggtcat
     4861 ggcctctcct gggaagagtc tccagaattt tgtaacagga ctttctctta gagatcagtg
     4921 tgttgtggcg acatgggaca atgtccaggc ttctgctgcc aga
//
