LOCUS       sequence-449787-        5032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0772m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical L6_IT_Car3 neurons 
            (note CLA neurons).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5032)
  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 5032)
  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: AiP13608 -
            pAAV-AiE0772m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      1044..1053
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1050..1052,1053..1055,1056..1766)
                     /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(1095..1116)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1356..1375)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1703..1724
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1859..1879)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2066..2177
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2295..2435
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2295..2424
                     /label=AAV2 ITR
     rep_origin      2510..2965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2597..2616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2807..2828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2982..3001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3101..3123
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3161..3179)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3247..3351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3352..3420,3421..4212)
                     /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(3570..3589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4383..4971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4872..4891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcccca ctcctgcctc tgcctataag
      181 agtttggcac agttggagat ttgtatacac aaatgcatgc gtggaaatac tgcgctctga
      241 tgacattcct cagggggcga cctttgatga tgaattttct tgcccttttt aggggaggag
      301 aagaggaggc gggtggtact tcacaagact aaatttactc aacttttttg ggtctaaatt
      361 tctttaggaa aaaaaatgtc aaatgattgt agatcccaaa ctgtctttgt tggcatttga
      421 ccccactcct gcctctgcct ataagagttt ggcacagttg gagatttgta tacacaaatg
      481 catgcgtgga aatactgcgc tctgatgaca ttcctcaggg ggcgaccttt gatgatgaat
      541 tttcttgccc tttttagggg aggagaagag gaggcgggtg gtacttcaca agactaaatt
      601 tactcaactt ttttgggtct aaatttcttt aggaaaaaaa atgtcaaatg attgtagatc
      661 ccaaactgtc tttgttggca tttgacccca ctcctgcctc tgcctataag agtttggcac
      721 agttggagat ttgtatacac aaatgcatgc gtggaaatac tgcgctctga tgacattcct
      781 cagggggcga cctttgatga tgaattttct tgcccttttt aggggaggag aagaggaggc
      841 gggtggtact tcacaagact aaatttactc aacttttttg ggtctaaatt tctttaggaa
      901 aaaaaatgtc aaatgattgt agatcccaaa ctgtctttgt tggcatttga gagctcgggc
      961 tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc
     1021 tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     1081 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1141 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
     1201 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt
     1261 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1321 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1381 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1441 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1501 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1561 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1621 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag
     1681 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1741 ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg cgaattcgat
     1801 atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1861 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1921 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg
     1981 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2041 attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2101 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2161 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2221 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg
     2281 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2341 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2401 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2461 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2521 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2581 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2641 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2701 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2761 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2821 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     2881 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2941 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3001 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3061 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3121 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3181 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3841 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3901 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3961 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4021 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4081 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4141 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4201 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4501 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
