LOCUS       sequence-470801-        4923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0058m_3xC2 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 4923)
  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 4923)
  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: AiP12021 -
            pAAV-AiE0058m_3xC2-minCMV-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..4923
                     /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        835..873
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      935..944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(941..943,944..946,947..1657)
                     /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(986..1007)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1247..1266)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1594..1615
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1750..1770)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1957..2068
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2186..2326
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2186..2315
                     /label=AAV2 ITR
     rep_origin      2401..2856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2488..2507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2698..2719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2873..2892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2992..3014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3052..3070)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3138..3242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3243..3311,3312..4103)
                     /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(3461..3480)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4274..4862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4763..4782
                     /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 gcgttcgttg ggacatcaaa ggatgttata ctgaagagat
      181 ctcaaagacc aggcagtcca gcctactcgt ttcccagaca agagaagtga agccaagaaa
      241 cagatgggct gcctcctcta gtcatatggg gcttaaaaat atatatctct atttttcatg
      301 agttttcatg aaagtgacca gtgttatctc atttcctgtc ttctcaaatt caggatccta
      361 cccaatctaa ctcgttggga catcaaagga tgttatactg aagagatctc aaagaccagg
      421 cagtccagcc tactcgtttc ccagacaaga gaagtgaagc caagaaacag atgggctgcc
      481 tcctctagtc atatggggct taaaaatata tatctctatt tttcatgagt tttcatgaaa
      541 gtgaccagtg ttatctcatt tcctgtcttc tcaaattcag gatcctaccc aatctaactc
      601 gttgggacat caaaggatgt tatactgaag agatctcaaa gaccaggcag tccagcctac
      661 tcgtttccca gacaagagaa gtgaagccaa gaaacagatg ggctgcctcc tctagtcata
      721 tggggcttaa aaatatatat ctctattttt catgagtttt catgaaagtg accagtgtta
      781 tctcatttcc tgtcttctca aattcaggat cctacccaat ctaacgagct cagaggtagg
      841 cgtgtacggt gggaggccta tataagcaga gctggtttag tgaaccgtca gatcgcctgg
      901 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
      961 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1021 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     1081 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     1141 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1201 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1261 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1321 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1381 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1441 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     1501 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     1561 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1621 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     1681 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1741 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1801 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     1861 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     1921 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     1981 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2041 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2101 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     2161 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2221 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2281 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2341 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2401 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2461 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2521 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2581 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2641 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2701 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     2761 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     2821 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     2881 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     2941 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3001 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3061 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3181 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3241 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3301 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3361 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3421 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3481 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3541 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3601 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3661 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3721 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3781 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3841 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3901 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3961 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4021 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4081 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4141 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4201 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4261 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4321 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4381 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4441 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4501 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4561 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4621 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4681 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4741 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4801 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4861 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4921 tgt
//
