LOCUS       sequence-466277-        5162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2281m 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 5162)
  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 5162)
  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: AiP1614 - pAAV-AiE2281m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      475..484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(481..483,484..486,487..1197)
                     /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(526..547)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(787..806)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1134..1155
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1290..1310)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1497..1608
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1726..1866
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1726..1855
                     /label=AAV2 ITR
     rep_origin      1941..2396
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2028..2047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2238..2259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2413..2432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2532..2554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2592..2610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2678..2782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2783..2851,2852..3643)
                     /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(3001..3020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3814..4402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4303..4322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4464..4593
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcgaggggcg cgcgctccgg gctgccgcag gtggcggcgc ggtgagcccg ggccaggtgc
       61 cccggcagcg gggcggcgct gtcgtgcggg acagccgggc tgccaggggc tcggagccgg
      121 gtcggagccc gcggggggcg gggagtgtgg cgagaaatgg ggaacaatgc gagtgagcaa
      181 cttgaggaag tcattgtgaa agaaagctgg gaattgctcc gcagccaact tagcagggca
      241 ctctaacaag tgcctgcgcg gccgcgcccg ggccggggac aggggcagcc cggcgcagta
      301 cagcccatcc cgggacgctc ggccgcggct gccggagacc cggtaggtcc cgcggggtgc
      361 aggagccccc agacgccggc ggagctcggg ctgggcataa aagtcagggc agagccatct
      421 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
      481 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      541 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      601 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      661 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      721 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      781 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      841 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      901 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
      961 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1021 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1081 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1141 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1201 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1261 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1321 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1381 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1441 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     1501 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1561 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1621 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1681 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     1741 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     1801 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     1861 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1921 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1981 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2041 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2101 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2161 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2221 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2281 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     2341 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2401 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2461 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2521 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2581 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2641 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2701 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2761 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2821 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2881 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2941 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3001 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3061 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3121 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3181 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3241 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3301 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3361 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3421 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3481 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3541 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3601 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3661 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3721 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3781 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3841 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3901 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3961 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4021 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4081 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4141 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4201 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4261 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4321 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4381 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4441 ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc
     4501 cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     4561 agggagtggc caactccatc actaggggtt cctgcggccg cacgcgttgt aagagtaata
     4621 atcgccggcg gccggccccg cacgtgccgc acccctgcgg gtcgagcgcg ggtcccgcag
     4681 tggcgcggtt gagccggacc ggcccgcggg caaagcgcgg gccgccgcat cccgcgcggg
     4741 gcagcggcgt gatgctgcgc tggaatgagg aagcttggcg ccaaggggag ggcccgggcg
     4801 cgctgcgcgc gggggtggcg gcggcgcgcc gagcgggccc ggcgcgggcg agcgggctgc
     4861 agccgccggc ggcgccagca ggtacggccg gtgcccgccg ccgcccggct accctcggcg
     4921 gccgggcccg agccccgtgc ggctgcaaac tttgtgtgcg cggcccggcg ggtcgcggag
     4981 ttgccgtctg aaggcgggac ccgagggcgg gcgcgggacg ggagggtgcc tccctctgac
     5041 ccgctcagtg tccccgtcag ggtcccgaag ccgcgtgccg tgcgcgcagg ccgggtgggc
     5101 tgtgggacaa gccgagcggg aggcgagtcg caagcgccaa cccaaagttt gcacggtgcg
     5161 gg
//
