LOCUS       sequence-470825-        5066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0460m_3xC3 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 5066)
  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 5066)
  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: AiP13025 -
            pAAV-AiE0460m_3xC3-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5066
                     /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      1078..1087
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1084..1086,1087..1089,1090..1800)
                     /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(1129..1150)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1390..1409)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1737..1758
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1893..1913)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2100..2211
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2329..2469
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2329..2458
                     /label=AAV2 ITR
     rep_origin      2544..2999
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2631..2650)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2841..2862
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3016..3035)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3135..3157
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3195..3213)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3281..3385
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3386..3454,3455..4246)
                     /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(3604..3623)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4417..5005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4906..4925
                     /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 gcgtaatgaa gctaaaaaca atctaacaac catcagggtg
      181 ctacagcagc acagaaatga ctagctcttt tgtgtcaagt tattgccggt tgtcttagtc
      241 actgtccgac tgatgtgcag agacaccatg actaaggcaa cgcttatcaa agaaagagtc
      301 tagttgagag cttgcttgta gttccagagg tttgtttagt ctagcatggt ggcacacact
      361 gtgctagaac aatattgaga gctacaacct gatctgaaaa gcagagagag atagagacag
      421 aaagaatgaa gctaaaaaca atctaacaac catcagggtg ctacagcagc acagaaatga
      481 ctagctcttt tgtgtcaagt tattgccggt tgtcttagtc actgtccgac tgatgtgcag
      541 agacaccatg actaaggcaa cgcttatcaa agaaagagtc tagttgagag cttgcttgta
      601 gttccagagg tttgtttagt ctagcatggt ggcacacact gtgctagaac aatattgaga
      661 gctacaacct gatctgaaaa gcagagagag atagagacag aaagaatgaa gctaaaaaca
      721 atctaacaac catcagggtg ctacagcagc acagaaatga ctagctcttt tgtgtcaagt
      781 tattgccggt tgtcttagtc actgtccgac tgatgtgcag agacaccatg actaaggcaa
      841 cgcttatcaa agaaagagtc tagttgagag cttgcttgta gttccagagg tttgtttagt
      901 ctagcatggt ggcacacact gtgctagaac aatattgaga gctacaacct gatctgaaaa
      961 gcagagagag atagagacag aaaggagctc gggctgggca taaaagtcag ggcagagcca
     1021 tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc
     1081 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1141 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1201 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     1261 accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     1321 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1381 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1441 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1501 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     1561 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1621 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1681 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1741 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1801 taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     1861 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1921 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     1981 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     2041 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     2101 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2161 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2221 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2281 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2341 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2401 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2461 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2521 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2581 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2641 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2701 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2761 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2821 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2881 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     2941 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3001 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3061 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3121 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3181 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3241 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3301 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3361 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3421 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3481 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3541 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3601 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3661 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3721 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3781 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3841 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3901 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3961 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4021 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4081 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4141 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4201 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4261 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4321 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4381 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4441 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4501 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5041 cttttgctgg ccttttgctc acatgt
//
