LOCUS       sequence-466259-        4630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2140m 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 4630)
  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 4630)
  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: AiP1429 - pAAV-AiE2140m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(528..1319,1320..1388))
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1389..1493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1561..1579
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1617..1639)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1739..1758
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1775..2230)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1912..1933)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2124..2143
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2305..2445
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2316..2445
                     /label=AAV2 ITR
     polyA_signal    2563..2674
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2861..2881
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2974..3684,3685..3687,3688..3690))
                     /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(3016..3037)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3365..3384
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3624..3645
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3687..3696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4208..4337
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4399..4630,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4479..4498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
       61 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1501 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     1561 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     1621 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     1681 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     1741 gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta
     1801 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc
     1861 aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg
     1921 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     1981 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     2041 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2101 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg
     2161 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2221 cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa
     2281 ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg
     2341 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     2401 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc
     2461 gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     2521 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     2581 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     2641 caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag ccacggaatt
     2701 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     2761 cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag ccatacggga
     2821 agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac
     2881 atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt gattatgata
     2941 tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca tgccgagagt
     3001 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     3061 tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc
     3121 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt
     3181 gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac
     3241 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     3301 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     3361 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     3421 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca
     3481 ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     3541 ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     3601 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     3661 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag
     3721 atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag
     3781 cccgagctcc gccggcgtgg gccatcactg agatttcata atgggaaaaa gtcacttcct
     3841 gatcttcttt ggtgcagaga ggaagaacgg tatgtttcaa tgtcccctgc ttaaaaaaaa
     3901 aaaagtggga aggttggctc attttcattg agaagaggaa actgaaaact ccagaggtta
     3961 aacaaatagg aaggtttatt tatttattta tttattttac aagccacaga gccagttaat
     4021 gattcctaag tcccaagtgg gtgttacatg tttgcttgaa ggctgggcca tgtgacatcc
     4081 caggcttggg gactctcctt cattctggct ccaggactcc agctccacga tgcgtcctgg
     4141 cctctgggag acatgcttct ggttgtgggg cgccggcgat tattactctt acaacgcgtg
     4201 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4261 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     4321 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4381 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4441 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4501 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4561 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4621 ggtgtaggtc
//
