LOCUS       sequence-479080-        4739 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  AiE3099m 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 4739)
  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 4739)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 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: AiP20073 - pAAV-AiE3099m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4739
                     /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      751..760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(757..759,760..762,763..1473)
                     /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(802..823)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1063..1082)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1410..1431
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1566..1586)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1773..1884
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2002..2142
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2002..2131
                     /label=AAV2 ITR
     rep_origin      2217..2672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2304..2323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2514..2535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2689..2708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2808..2830
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2868..2886)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2954..3058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3059..3127,3128..3919)
                     /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(3277..3296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4090..4678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4579..4598
                     /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 gcgtgaaata ggtgaccttt tatgtggaca ataaagcccc
      181 ccttccttaa tcaagatctc tcagaggaaa cacagtggtg tccattcaga ttgctttctt
      241 ccgtatgtcc tggtggagcg attctgccat gataggcagc aaggtcagcc tgagtgctcc
      301 tgctcctctt ttcatcccgt tgtgttgagt ttgctctcac agggtgttag atgctgctct
      361 aagaaatagc tctcactgtc tcccttctct tcatcaactt cctcagttgg ggatgagagg
      421 taatataatt ctgagtaacc aggcccctct tgaaatctca tttcagcccc aaaccggctc
      481 agttcccagc caagagcaga gcacctggca gacagaatta cctcatagta atgtaagaag
      541 caggactgtc atcattcacg gttaactctg tgtttccact gtccctagac tggcctgttc
      601 tcttccattt ttgcttctgt ccttgacaca aatgaaaaaa aataccttgc gccggcggag
      661 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
      721 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
      781 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      841 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
      901 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
      961 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1021 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1081 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1141 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1201 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     1261 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     1321 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     1381 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1441 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     1501 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     1561 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     1621 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     1681 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1741 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     1801 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1861 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1921 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     1981 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2041 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2101 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2161 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2221 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2281 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2341 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2401 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2461 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2521 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     2581 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2641 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     2701 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     2761 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2821 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     2881 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     2941 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3001 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3061 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4021 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4081 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4201 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
