LOCUS       sequence-460606-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0121h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  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 5126)
  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: AiP12786 -
            pAAV-AiE0121h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5126
                     /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      1138..1147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1144..1146,1147..1149,1150..1860)
                     /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(1189..1210)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1450..1469)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1797..1818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1953..1973)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2160..2271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2389..2529
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2389..2518
                     /label=AAV2 ITR
     rep_origin      2604..3059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2691..2710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2901..2922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3076..3095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3195..3217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3255..3273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3341..3445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3446..3514,3515..4306)
                     /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(3664..3683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4477..5065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4966..4985
                     /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 gcgtctatct ggtttataat tagaagtgaa atgattttaa
      181 atggcttcca catggcttag ttttccaagc agttccagtt acatagaact tcatataaac
      241 agataatgag aggctagagt tggagagcac agttcctgct tccaaattgg cttgaagtgg
      301 ggacatctgc tctgcatgaa gcctgggtca ggctgcccct ctcatctgtg acctcctgtg
      361 gtcgggaggg acttgctctg gattatataa aggcttcaca agagtactgg aagcggggac
      421 ggaatcaact tgagcaaaaa acctctatct ggtttataat tagaagtgaa atgattttaa
      481 atggcttcca catggcttag ttttccaagc agttccagtt acatagaact tcatataaac
      541 agataatgag aggctagagt tggagagcac agttcctgct tccaaattgg cttgaagtgg
      601 ggacatctgc tctgcatgaa gcctgggtca ggctgcccct ctcatctgtg acctcctgtg
      661 gtcgggaggg acttgctctg gattatataa aggcttcaca agagtactgg aagcggggac
      721 ggaatcaact tgagcaaaaa acctctatct ggtttataat tagaagtgaa atgattttaa
      781 atggcttcca catggcttag ttttccaagc agttccagtt acatagaact tcatataaac
      841 agataatgag aggctagagt tggagagcac agttcctgct tccaaattgg cttgaagtgg
      901 ggacatctgc tctgcatgaa gcctgggtca ggctgcccct ctcatctgtg acctcctgtg
      961 gtcgggaggg acttgctctg gattatataa aggcttcaca agagtactgg aagcggggac
     1021 ggaatcaact tgagcaaaaa acctgagctc gggctgggca taaaagtcag ggcagagcca
     1081 tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc
     1141 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1201 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1261 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     1321 accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     1381 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1441 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1501 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1561 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     1621 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1681 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1741 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1801 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1861 taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     1921 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1981 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2041 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     2101 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     2161 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2221 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2281 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2341 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2401 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2461 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2521 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2581 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2941 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3001 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3061 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3121 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3181 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3241 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3841 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3961 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4021 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4081 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4141 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4201 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4261 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4321 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4381 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4441 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4501 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4561 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4621 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4681 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4741 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4801 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4861 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4921 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4981 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5041 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5101 cttttgctgg ccttttgctc acatgt
//
