LOCUS       sequence-449767-        4830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0770m 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 4830)
  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 4830)
  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: AiP12603 - pAAV-AiE0770m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4830
                     /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      842..851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(848..850,851..853,854..1564)
                     /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(893..914)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1154..1173)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1501..1522
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1657..1677)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1864..1975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2093..2233
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2093..2222
                     /label=AAV2 ITR
     rep_origin      2308..2763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2395..2414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2605..2626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2780..2799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2899..2921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2959..2977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3045..3149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3150..3218,3219..4010)
                     /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(3368..3387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4181..4769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4670..4689
                     /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 cgcgtatggg aaagcagcac gtgttcttaa
      181 ctgctagact cctatttacc atgacagtaa tatcattatt atttttaaat cattttataa
      241 tttcaggaga aagcaaatga ttgctaataa cggtctccct cttcagtaca tatgcactac
      301 acgcgcatgt gtcctttaaa gataaatttg agttggaagc atcggtatga gctggtgagt
      361 tttaagaagt tccagaaaaa aaaaagaagt tccagtattt cctagatttg tcctctgcaa
      421 agtcctaggc ataaaggcag cgcagtttgt ggcctttaaa taccacttcc cacgaaagca
      481 accaaggctc cttggagacg tgagcgattt gatgtctata aatatgagca cacagtgctg
      541 tgcttcccca ctgaggtcag aaagaccact accctgaatc caaaccacct tcatggctta
      601 gcagatgagg ccagagctgg acaagagcaa ttaaagggaa caattacatt acatgttggt
      661 gtttgattta tctgcagatt tttttaacca gtaaagttta ttttgaaatg caattaattt
      721 gtaagatcag accgcccatt ggttagttga gctcgggctg ggcataaaag tcagggcaga
      781 gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt
      841 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      901 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      961 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
     1021 gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca
     1081 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1141 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1201 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1261 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
     1321 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     1381 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1441 caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     1501 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1561 caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     1621 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1681 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     1741 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     1801 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     1861 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1921 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1981 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2041 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     2101 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2161 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2221 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2281 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     2341 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2401 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2461 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2521 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2581 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2641 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2701 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2761 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     2821 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2881 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2941 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3001 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3241 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3301 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3361 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3421 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3541 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3661 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3721 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4021 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4081 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4141 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4261 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4321 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4801 tggccttttg ctggcctttt gctcacatgt
//
