LOCUS       sequence-449741-        4946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2016m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical L5_IT neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  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 4946)
  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: AiP14477 -
            pAAV-AiE2016m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4946
                     /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      958..967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(964..966,967..969,970..1680)
                     /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(1009..1030)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1270..1289)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1617..1638
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1773..1793)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1980..2091
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2209..2349
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2209..2338
                     /label=AAV2 ITR
     rep_origin      2424..2879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2511..2530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2721..2742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2896..2915)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3015..3037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3075..3093)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3161..3265
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3266..3334,3335..4126)
                     /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(3484..3503)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4297..4885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4786..4805
                     /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 gcgtctgtgc tctgacagcc gagggtcaga gctggaatca
      181 cactcttgtt gtcttttaat ctctccctcc ctttcttcct tctttcttca ctctgttgtg
      241 attgctcatg gaacagatcc tagctggtct ccctggcaac ctacatgatt tgagcccaac
      301 agatggataa tggggacatc gacttccaat gtcattcaac agaatcattg ccaagggagt
      361 ctgatgagca ggcaactgag atgactgtgc tctgacagcc gagggtcaga gctggaatca
      421 cactcttgtt gtcttttaat ctctccctcc ctttcttcct tctttcttca ctctgttgtg
      481 attgctcatg gaacagatcc tagctggtct ccctggcaac ctacatgatt tgagcccaac
      541 agatggataa tggggacatc gacttccaat gtcattcaac agaatcattg ccaagggagt
      601 ctgatgagca ggcaactgag atgactgtgc tctgacagcc gagggtcaga gctggaatca
      661 cactcttgtt gtcttttaat ctctccctcc ctttcttcct tctttcttca ctctgttgtg
      721 attgctcatg gaacagatcc tagctggtct ccctggcaac ctacatgatt tgagcccaac
      781 agatggataa tggggacatc gacttccaat gtcattcaac agaatcattg ccaagggagt
      841 ctgatgagca ggcaactgag atgagagctc gggctgggca taaaagtcag ggcagagcca
      901 tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc
      961 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1021 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1081 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     1141 accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     1201 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1261 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1321 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1381 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     1441 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1501 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1561 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1621 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1681 taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     1741 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1801 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     1861 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     1921 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     1981 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2041 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2101 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2161 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2221 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2281 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2341 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2401 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2701 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2761 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     2821 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     2881 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     2941 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3001 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3061 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3121 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3181 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3241 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3301 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3361 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3421 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3481 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3541 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3601 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3661 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3721 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3781 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3841 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3901 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3961 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4021 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4081 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4141 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4201 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4261 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4321 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4381 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4441 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4501 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4561 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4621 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4681 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4741 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4801 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4861 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4921 cttttgctgg ccttttgctc acatgt
//
