LOCUS       sequence-450564-        5123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0049m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical L4/5_IT neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5123)
  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 5123)
  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: AiP12579 - pAAV-AiE0049m-minCMV-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..5123
                     /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        1035..1073
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1135..1144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1141..1143,1144..1146,1147..1857)
                     /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(1186..1207)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1447..1466)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1794..1815
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1950..1970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2157..2268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2386..2526
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2386..2515
                     /label=AAV2 ITR
     rep_origin      2601..3056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2688..2707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2898..2919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3073..3092)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3192..3214
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3252..3270)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3338..3442
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3443..3511,3512..4303)
                     /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(3661..3680)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4474..5062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4963..4982
                     /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 gcgttaggtg ctgtcattta cctacggttg tctccaaatt
      181 tcttcaacca agtagagaaa aatgagagag aaggaaagaa aaaaagaggt atggggagaa
      241 gagaaagaag gcaacttgtt aaaaatctca gtcaaactta catactatat agaacagcat
      301 ggtgaattta gggcacatgg atataaaatg gaagtttctt attcagtagc agcaacttgt
      361 gggcacagga gttggcaaag ataaaaatgt ccaaagtcac aaatacaatg tatagttagt
      421 cataggtgct gttatttgcc tcaaaaaata gacttttatt ttgcctttct tttctttaac
      481 cacactcaaa attagagaac agagacaaaa cccagcagga aatagcacag aaagcccaca
      541 gaatcaaaga cgtgttcaaa cagccagctg aattcattgc acatttcaac cacagaaata
      601 ttttcaggtg attctgttgt ttgacaaaac gtgggaacca caggatctac aacacttgca
      661 agcaaaactc aacagctcta ataatagtta cagaagtgaa agccaatttg gataaaataa
      721 gacattgact caagtcctct cagaagagtt ttgaaagcaa agtttacaaa agtctggttt
      781 gtcctttggg atttacagac ctttcagccc cttgattcat tttttttttt ttggatttct
      841 tcatcactgg gagaattccc atgcattatt tctcccctgc ttcaaaatca tcaaatgtga
      901 aacatttttc actcttttct tctgtatata gtgataaaat agctattggc ttttggctaa
      961 atgtgctact ttgagcccac ccacacaagg gagaaatggg ggcagacatg agtttgggca
     1021 tgagtgagct cagaggtagg cgtgtacggt gggaggccta tataagcaga gctggtttag
     1081 tgaaccgtca gatcgcctgg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
     1141 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1201 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1261 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1321 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     1381 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1441 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1501 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1561 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     1621 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1681 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1741 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1801 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1861 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1921 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1981 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2041 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2101 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     2161 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2221 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2281 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2341 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2401 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2461 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2521 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2581 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2941 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     3001 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3061 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3121 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3181 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3241 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3301 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3361 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3421 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3481 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3541 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3601 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3661 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3721 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3781 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3841 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3901 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3961 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4021 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4081 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4141 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4201 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4261 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4321 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4381 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4441 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4501 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4561 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4621 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4681 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4741 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4801 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4861 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4921 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4981 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5041 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5101 ttgctggcct tttgctcaca tgt
//
