LOCUS       sequence-462000-        4787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2401m 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 4787)
  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 4787)
  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: AiP1730 - pAAV-AiE2401m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      799..808
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(805..807,808..810,811..1521)
                     /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(850..871)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1111..1130)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1458..1479
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1614..1634)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1821..1932
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2050..2190
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2050..2179
                     /label=AAV2 ITR
     rep_origin      2265..2720
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2352..2371)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2562..2583
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2737..2756)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2856..2878
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2916..2934)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3002..3106
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3107..3175,3176..3967)
                     /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(3325..3344)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4138..4726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4627..4646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaag agtaataatc ggcaaaaaac ttcatctttc
      181 atgagtgcaa tttcaaactt ggtgccaacg agagacatct tccctggact caaatgcgcg
      241 aagcctgaac aggacatctt gagtgttagc ttctctggga tggctgcggt gtgcagcaca
      301 tttttgttaa ggtgttattc aaagtttctg attaacataa gtttttctta aaattataca
      361 aaaacataat accatgatta tggaaataaa aagccaccca caagcttact atctttacaa
      421 accatctatt ttcattcttt cctgtctttt ccaaaaaagt attacctcct cccgcccctc
      481 agtgttggac acacagcaca aatatgcctt tgtgttctga tcttttttgt gtgcctcaca
      541 caatgagtaa tatcagtgtg ctcttaagaa taacagattc aaataacttc caagcattac
      601 attatttgat acattctcct ttatatcagc atccagatat cacggtggga ctggtaggtt
      661 agctggcatt cacttttgtt tccagtaata gcttgttttt ccccggagct cgggctgggc
      721 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
      781 aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      841 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      901 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
      961 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
     1021 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1081 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1141 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1201 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1261 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1321 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1381 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     1441 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1501 ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat
     1561 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1621 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     1681 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     1741 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1801 gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1861 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1921 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1981 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     2041 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2101 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2161 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     2221 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     2281 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2341 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2401 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2461 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2521 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2581 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2641 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2701 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     2761 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2821 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2881 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2941 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3001 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3061 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3121 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3181 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3241 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3301 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3361 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3421 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3481 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3541 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3601 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3661 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3721 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3781 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3841 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3901 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3961 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4021 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4081 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4141 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4201 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4261 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4321 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4381 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4441 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4501 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4561 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4621 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4681 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4741 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
