LOCUS       sequence-449736-        4913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0774m_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 4913)
  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 4913)
  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: AiP13041 -
            pAAV-AiE0774m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4913
                     /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      925..934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(931..933,934..936,937..1647)
                     /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(976..997)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1237..1256)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1584..1605
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1740..1760)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1947..2058
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2176..2316
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2176..2305
                     /label=AAV2 ITR
     rep_origin      2391..2846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2478..2497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2688..2709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2863..2882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2982..3004
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3042..3060)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3128..3232
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3233..3301,3302..4093)
                     /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(3451..3470)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4264..4852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4753..4772
                     /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 gcgtacagca agtgttctat gattttcaac aggacacttg
      181 gatctagcca actcagtttt caagcctgcc cccgagttgt agatcctgtc ttcacattct
      241 aaactaatct cacaccactt tgtgccaact ccactctggt aaagaccatg cactcgagtt
      301 agtgcaagac attttgttct ggcctcgggg tgtgctgccg cctcgtcagg atgggaggtg
      361 gaggcgagtc cagacagcaa gtgttctatg attttcaaca ggacacttgg atctagccaa
      421 ctcagttttc aagcctgccc ccgagttgta gatcctgtct tcacattcta aactaatctc
      481 acaccacttt gtgccaactc cactctggta aagaccatgc actcgagtta gtgcaagaca
      541 ttttgttctg gcctcggggt gtgctgccgc ctcgtcagga tgggaggtgg aggcgagtcc
      601 agacagcaag tgttctatga ttttcaacag gacacttgga tctagccaac tcagttttca
      661 agcctgcccc cgagttgtag atcctgtctt cacattctaa actaatctca caccactttg
      721 tgccaactcc actctggtaa agaccatgca ctcgagttag tgcaagacat tttgttctgg
      781 cctcggggtg tgctgccgcc tcgtcaggat gggaggtgga ggcgagtcca ggagctcggg
      841 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      901 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      961 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1021 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1081 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1141 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1201 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1261 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1321 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1381 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1441 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1501 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1561 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1621 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     1681 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1741 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1801 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     1861 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1921 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     1981 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2041 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2101 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     2161 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2221 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2281 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2341 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2401 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2461 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2521 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2581 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2641 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2701 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     2761 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2821 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     2881 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2941 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3001 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3061 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3121 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3181 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3241 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3301 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3361 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3421 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3481 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3541 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3601 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3661 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3721 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3781 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3841 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3901 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3961 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4021 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4081 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4141 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4381 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
