LOCUS       sequence-462009-        4836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2573m 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 4836)
  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 4836)
  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: AiP1886 - pAAV-AiE2573m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        602..614
                     /gene="Pichia pastoris ADE2"
                     /label=PpADE2 HC promoter
                     /note="weak 13-bp promoter fragment that requires high-copy
                     integration for efficient expression"
     regulatory      848..857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(854..856,857..859,860..1570)
                     /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(899..920)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1160..1179)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1507..1528
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1663..1683)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1870..1981
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2099..2239
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2099..2228
                     /label=AAV2 ITR
     rep_origin      2314..2769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2401..2420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2611..2632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2786..2805)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2905..2927
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2965..2983)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3051..3155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3156..3224,3225..4016)
                     /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(3374..3393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4187..4775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4676..4695
                     /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 gcggccgcat ccccacagct cagtctaatg tttgactgaa agtctttgaa
      181 tctgttcaag tcagctgcta ggtgaagcct ctcagaggac agtatgctag gctcctgtct
      241 gcaaatgcat taacaatgtc agggattggt tctagcccat gggatggatc tcaaactgag
      301 tcagtcattg gctggccatt ccctcaatct ctgctccatc tttgtccttc cacatcttgt
      361 aggcaacaca aatttggggt ggaagttttt gtgggtgggg tgctgtcttt atccctctgc
      421 tgggagtcct gccagtttac agattacagt ttacagttca cttcaagctc catatgtccc
      481 actgctagga atctcagcta gattcattcc cagagattcc ccaaagcctc ccccactcca
      541 gatctccagc acatcctaga gatgctccct ccccctacaa gcaacttttt tttcattctc
      601 tcttccaaac tctccctaca cagatctgcc cctgactgtt tcctggccca tcctctctcc
      661 catacagttc cctccctcca ttcacctcag atgtctattt tatttcccct tctgcataag
      721 attctagcat cctcccttgg gtccttcttg ttcggagctc gggctgggca taaaagtcag
      781 ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct
      841 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      901 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      961 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
     1021 gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc
     1081 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1141 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1201 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1261 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
     1321 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
     1381 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1441 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
     1501 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1561 gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct
     1621 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1681 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     1741 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     1801 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga
     1861 gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1921 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1981 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2041 gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag
     2101 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2161 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     2221 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2281 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     2341 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     2401 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2461 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2521 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2581 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2641 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     2701 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2761 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2821 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2881 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2941 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3001 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3061 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3121 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3181 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3241 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3301 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3361 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3421 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3481 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3541 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3601 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3661 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3721 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3781 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3841 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3901 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3961 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4021 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4081 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4141 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4201 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4261 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4321 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4381 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4441 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4501 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4561 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4621 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4681 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4741 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4801 ggttcctggc cttttgctgg ccttttgctc acatgt
//
