LOCUS       sequence-449739-        5216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2182m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical L5_IT neurons 
            (strong).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5216)
  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 5216)
  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: AiP14467 -
            pAAV-AiE2182m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1228..1237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1234..1236,1237..1239,1240..1950)
                     /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(1279..1300)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1540..1559)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1887..1908
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2043..2063)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2250..2361
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2479..2619
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2479..2608
                     /label=AAV2 ITR
     rep_origin      2694..3149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2781..2800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2991..3012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3166..3185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3285..3307
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3345..3363)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3431..3535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3536..3604,3605..4396)
                     /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(3754..3773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4567..5155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5056..5075
                     /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 gcgtggcaag tgacatttat tgacagcact gtgaactaca
      181 tatagttctg taagccgcct gctgccacac taactgggtt tctgagtggg aggccagagc
      241 tgtatggaag gaaacggcga gagaaataat ggaggccaac atatgtttct gttcaagatc
      301 cccaaagttt actaagagtc tgtgcttata aagggggagg cccatccccc gccaatccat
      361 tcttggtgcc tgttgccagc ctgtaggcat atctgctgga tgctgtctca ggaatatctc
      421 aagggcctct cagcaggttg cagtgtcttg gagaagagca gcagtagttg acatggcaag
      481 tgacatttat tgacagcact gtgaactaca tatagttctg taagccgcct gctgccacac
      541 taactgggtt tctgagtggg aggccagagc tgtatggaag gaaacggcga gagaaataat
      601 ggaggccaac atatgtttct gttcaagatc cccaaagttt actaagagtc tgtgcttata
      661 aagggggagg cccatccccc gccaatccat tcttggtgcc tgttgccagc ctgtaggcat
      721 atctgctgga tgctgtctca ggaatatctc aagggcctct cagcaggttg cagtgtcttg
      781 gagaagagca gcagtagttg acatggcaag tgacatttat tgacagcact gtgaactaca
      841 tatagttctg taagccgcct gctgccacac taactgggtt tctgagtggg aggccagagc
      901 tgtatggaag gaaacggcga gagaaataat ggaggccaac atatgtttct gttcaagatc
      961 cccaaagttt actaagagtc tgtgcttata aagggggagg cccatccccc gccaatccat
     1021 tcttggtgcc tgttgccagc ctgtaggcat atctgctgga tgctgtctca ggaatatctc
     1081 aagggcctct cagcaggttg cagtgtcttg gagaagagca gcagtagttg acatgagctc
     1141 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
     1201 gatctttcga agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
     1261 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1321 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     1381 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
     1441 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1501 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1561 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1621 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1681 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1741 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1801 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     1861 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1921 atcactctcg gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt
     1981 cgatatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2041 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2101 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg
     2161 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2221 gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc catctgttgt
     2281 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2341 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2401 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg agatctcacg
     2461 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2521 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2581 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2641 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     2701 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2761 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2821 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2881 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     2941 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3001 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3061 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3121 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3181 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3241 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3301 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3361 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3421 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3481 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3541 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3601 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3661 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3721 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3781 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3841 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3901 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3961 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4021 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4081 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4141 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4201 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4261 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4321 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4381 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4441 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4681 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4801 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4861 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4921 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4981 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5041 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5101 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5161 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
