LOCUS       sequence-473568-        4519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0731m 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 4519)
  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 4519)
  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: AiP12758 - pAAV-AiE0731m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4519
                     /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      531..540
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(537..539,540..542,543..1253)
                     /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(582..603)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(843..862)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1190..1211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1346..1366)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1553..1664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1782..1922
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1782..1911
                     /label=AAV2 ITR
     rep_origin      1997..2452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2084..2103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2294..2315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2469..2488)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2588..2610
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2648..2666)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2734..2838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2839..2907,2908..3699)
                     /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(3057..3076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3870..4458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4359..4378
                     /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 gcgtagaagc tcttagtgcc aggtccccct catggataag
      181 gaaattgagg cagtgaatta aaccagatcc ctacgcaagc ccagccacct tactttatcg
      241 tttgtgaaac atcaccacac ttctttgctt cccaacacct cagtctcctc cctgtcacag
      301 cagcaacggg gagggaagga gatggatgag tggtttccca agacatcttg gtggcaagcc
      361 tcaaaataaa cctttctggg tagtcaaatc cagagaaacg agaagtgtag agaaggctga
      421 gaagaagtca tttcgcagag ctcgggctgg gcataaaagt cagggcagag ccatctattg
      481 cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc gccaccatgg
      541 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      601 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      661 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      721 tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
      781 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      841 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      901 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
      961 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     1021 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     1081 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1141 tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1201 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg
     1261 acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg
     1321 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1381 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1441 aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1501 caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct
     1561 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1621 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1681 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1741 aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     1801 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     1861 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     1921 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     1981 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2041 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     2101 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2161 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2221 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2281 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     2341 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     2401 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     2461 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     2521 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2581 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     2641 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2701 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2761 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2821 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2881 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2941 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3001 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3061 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3121 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3181 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3241 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3301 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3361 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3421 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3481 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3541 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3601 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3661 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3721 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3781 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3841 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4441 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4501 tggccttttg ctcacatgt
//
