LOCUS       sequence-433027-        4757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2004m 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 4757)
  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 4757)
  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: AiP1492 - pAAV-AiE2004m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     588..607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(826..930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     998..1016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1054..1076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1176..1195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1212..1667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1349..1370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1561..1580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1742..1882
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1753..1882
                     /label=AAV2 ITR
     polyA_signal    2000..2111
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2298..2318
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2383..3093,3094..3096,3097..3099))
                     /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(2425..2446)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2774..2793
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3033..3054
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3096..3105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3772..3901
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3963..4551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4043..4062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4722..4757,1..756,757..825))
                     /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 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
       61 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      121 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
      181 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
      241 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
      301 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
      361 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      421 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
      481 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
      541 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
      601 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
      661 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      721 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      781 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
      841 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      901 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
      961 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1021 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     1081 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     1141 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     1201 agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1261 aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga
     1321 atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1381 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1441 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1501 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1561 agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg
     1621 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt
     1681 tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1741 ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
     1801 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     1861 aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtgagatct catgcctgct
     1921 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     1981 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     2041 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     2101 agaaggcaca gtcgaagatc tctcgagcca cggaattgtc agtgcccaac agccgagccc
     2161 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaaga actaaccagg
     2221 atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca
     2281 ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc
     2341 tttcacaaat tttgtaatcc agaggttgat tatgatatct tacttgtata gctcgtccat
     2401 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     2461 gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac
     2521 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc
     2581 ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     2641 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     2701 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2761 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     2821 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     2881 ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
     2941 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
     3001 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     3061 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcggatc cgagctcgct
     3121 agcttcgaaa gatctggatc ccagaagcaa atgtaagcaa tagatggctc tgccctgact
     3181 tttatgccca gcccgagctc atttgtcctg tgcatagcat tgcctctttt cactaagttt
     3241 tctgtggcca gttgtgtctc cagggtggaa cagattgctg ttgctttagc tgagcaccag
     3301 atgaggtact tggcttgcat tgaggggctc atgttgaatg aaaattatgc ttgtctctat
     3361 ttttttggct ccgcttgtag aactgaatgt gggcagatta aatcaatgta catatgatac
     3421 aactcaccat aaaacacttg ctcaaacaat cttttcgtga gccaacagga atagacagga
     3481 ataaaaataa acaccggtgc cagattacag agttcctgat cagcacacgt tgataggaaa
     3541 ttaatgtgca tgtattgaaa tagcagaaga tgatgtgttc tgagatctct ctgaaagcaa
     3601 gttagcatgg caatgcaagt tagtgagtca gtgatattca acagggtaac gcttccaagt
     3661 gtgagtgttg aagcttttat aaaatgattg catttaaaat gtaatttgta gcaaatccaa
     3721 ggactcaatg taatagtttg attgccccca aggcttatgt ttcgcggccg caggaacccc
     3781 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3841 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3901 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4381 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4741 gaggcaccta tctcagc
//
