LOCUS       sequence-432534-        4780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2156m 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 4780)
  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 4780)
  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: AiP1385 - pAAV-AiE2156m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   228..368
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   239..368
                     /label=AAV2 ITR
     polyA_signal    486..597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     784..804
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(897..1607,1608..1610,1611..1613))
                     /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(939..960)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1288..1307
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1547..1568
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1610..1619
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2281..2410
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2472..3060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2552..2571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3231..4022,4023..4091))
                     /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     3854..3873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4092..4196)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4264..4282
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4320..4342)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4442..4461
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4478..4780,1..153))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4615..4636)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
       61 aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
      121 ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg
      181 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct
      241 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
      301 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
      361 gggttcctgc ggccgctcgg tccgcacgtg agatctcatg cctgctattg tcttcccaat
      421 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
      481 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
      541 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
      601 aagatctctc gagccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
      661 gcaaggcagg cggcgatgag ttccgccgtg gcaagaacta accaggattt atacaaggag
      721 gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat
      781 ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg
      841 taatccagag gttgattatg atatcgaatt cgcggccgcg gcgcgccgtc gacttacttg
      901 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
      961 tgatcgcgct tctcgttggg gtctttgctc agcttggact ggtagctcag gtagtggttg
     1021 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1081 tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc
     1141 tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     1201 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     1261 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1321 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     1381 tggtcggggt agcgggcgaa gcactgcacg ccgtagccca gggtggtcac gagggtgggc
     1441 cagggcacgg gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg
     1501 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     1561 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     1621 accggtagcg ctagcttcga aagatctgga tcccagaagc aaatgtaagc aatagatggc
     1681 tctgccctga cttttatgcc cagcccgagc tcctccaaag gactgccaat aacaagttca
     1741 ggggaagcac aaacaggtct cagcttccgt cttttcctct ttcctccaat gagattctct
     1801 agcaataatc agccaagaag agttactgta gggaagccgg aacataagcg tcaccgtcat
     1861 cagactacaa ctcaagcagc gcctaccgaa gaatcctctg tgaaaccaca ggcttcaggc
     1921 gggcatggtg gcatggggga gcgcattttc cctggcccac gttcccagca tgcttctcgg
     1981 gttggagaat gccattcaga tagagatggg agtgaactat acctctacaa aaccaatagg
     2041 aaagaaattt cagtgggaaa atcactaaga agcaatcaga agtaatgtgg ctaaatcagg
     2101 caacatgagt atacaatcaa acacgggcac atttagaaag tcctggtccc cagctaataa
     2161 aaccggggtc atgagaactc tctgcactac catctgcgtt aactggctct cccagccctg
     2221 tgccagcaga agaaaaacaa agaggaggag aggcagttct ttagcaacgc gtgcggccgc
     2281 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2341 ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2401 tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2461 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2521 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2581 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2641 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2701 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2761 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2821 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2881 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2941 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3001 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3061 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3121 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3181 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3241 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3301 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3361 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3421 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3481 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3541 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3601 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3661 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3721 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3781 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3841 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3901 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3961 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4021 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4081 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4141 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4201 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4261 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4321 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4381 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4441 gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg
     4501 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc
     4561 ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt
     4621 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4681 tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg
     4741 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
//
