LOCUS       sequence-466335-        5107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2497m 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 5107)
  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 5107)
  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: AiP1986 - pAAV-AiE2497m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(134..153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      947..1535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1436..1455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1597..1726
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2715..2724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2721..2723,2724..2726,2727..3437)
                     /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(2766..2787)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3027..3046)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3374..3395
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3530..3550)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3737..3848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3966..4106
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3966..4095
                     /label=AAV2 ITR
     rep_origin      4181..4636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4268..4287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4478..4499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4653..4672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4772..4794
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4832..4850)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4918..5022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5023..5091,5092..5107,1..776)
                     /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"
ORIGIN
        1 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
       61 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      121 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      181 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      241 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      301 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
      361 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
      421 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
      481 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      541 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      601 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
      661 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      721 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
      781 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      841 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      901 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      961 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1021 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1081 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1141 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1201 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1261 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1321 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1381 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1441 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1501 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1561 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     1621 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     1681 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacactg
     1741 taaacatatt ttgtttgatt gattaatatt atagatgtca ttgattttac tgctgaaatg
     1801 tttcctggtg tctctcatct tgcagaaatc atgtttatat gctgcttgag acttattcat
     1861 gggcttagta aagaaggttc aagtgagatg atagtaccta ttaattatct tatttgcttt
     1921 gttttaaact ttcatctttt tatgtaaata actattgagt agaaaaatgg ttcaaaaaat
     1981 ttcccaactc ttatataaat tactaatttg tcaccccaag gtttgctttg gagctgagtt
     2041 ttcttgcctt tggattacct gctggtcttt tcagctggat gatgtggatt ttcacagcct
     2101 attgcctttt caatagaata attgtgcctc tgccaagtgt ataatcatgt gttttccctt
     2161 atatcatcca catgtggaca gcagcctgcc aacacataaa caagaaccag gttacgtcac
     2221 aaacagatgc tgctcaaata ccaaagcacc gtgccctcct ctcgagttat gtttatttgt
     2281 gcacagtggg gattaatatt ttttttgtgt gtgaaagaag atagagttac ttttgctgtt
     2341 ttcaaatagc agaagtgaat gtcccatgca tgagttcatg aaatctgtgc ccacaaggcc
     2401 tgatacttta atagggatga tagacaaatt gtctgggaaa gttgagcttt gtaagtcgtg
     2461 ttggtattga aaggaggtag ggaaattgaa aggatgattc ttttttaaac tattgtctat
     2521 ctatctatct atctatctat ctatctatct atctatctat ctatctgtct gtctgtctgt
     2581 ctggtttgat gtgtgcctgt gttcgtgcat gtgcatgcac ggagctcggg ctgggcataa
     2641 aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc
     2701 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2761 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2821 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     2881 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
     2941 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3001 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3061 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3121 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3181 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     3241 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3301 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     3361 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3421 tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc
     3481 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3541 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3601 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     3661 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3721 ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3781 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3841 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3901 agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg
     3961 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4021 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4081 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4141 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     4261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4321 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4381 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4501 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4561 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4621 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4681 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4741 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4801 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4861 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5101 acgctgg
//
