LOCUS       sequence-470942-        4910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0170h_3xC1 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 4910)
  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 4910)
  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: AiP13562 -
            pAAV-AiE0170h_3xC1-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4910
                     /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      922..931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(928..930,931..933,934..1644)
                     /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(973..994)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1234..1253)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1581..1602
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1737..1757)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1944..2055
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2173..2313
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2173..2302
                     /label=AAV2 ITR
     rep_origin      2388..2843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2475..2494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2685..2706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2860..2879)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2979..3001
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3039..3057)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3125..3229
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3230..3298,3299..4090)
                     /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(3448..3467)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4261..4849
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4750..4769
                     /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 gcgtatttat tccatataca cataaatagc accaccatgc
      181 acctgtggcc aaaagccaga aaatggagtg ccatcctcca caacacctta ctcacactac
      241 cttgatgcct agatggtctt aaagtcttaa agattctgct tccttattac ttctaaaatc
      301 tgttcccacc accctacttc tactgtcttt tagctaatct cacaatattc aatatttctt
      361 gccaggatta ttatttattc catatacaca taaatagcac caccatgcac ctgtggccaa
      421 aagccagaaa atggagtgcc atcctccaca acaccttact cacactacct tgatgcctag
      481 atggtcttaa agtcttaaag attctgcttc cttattactt ctaaaatctg ttcccaccac
      541 cctacttcta ctgtctttta gctaatctca caatattcaa tatttcttgc caggattatt
      601 atttattcca tatacacata aatagcacca ccatgcacct gtggccaaaa gccagaaaat
      661 ggagtgccat cctccacaac accttactca cactaccttg atgcctagat ggtcttaaag
      721 tcttaaagat tctgcttcct tattacttct aaaatctgtt cccaccaccc tacttctact
      781 gtcttttagc taatctcaca atattcaata tttcttgcca ggattattga gctcgggctg
      841 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
      901 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      961 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1021 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1081 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1141 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1201 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1261 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1321 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1381 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1441 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1501 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     1561 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1621 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     1681 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1741 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1801 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     1861 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     1921 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1981 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2041 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2101 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     2161 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2221 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2281 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2341 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     2401 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2461 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2521 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2581 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     2641 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2701 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     2761 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2821 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     2881 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2941 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3001 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3061 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3121 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3181 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3241 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3301 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3361 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3421 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3481 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3541 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3601 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3661 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3721 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3781 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3841 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3901 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3961 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4021 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4081 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4141 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4201 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4261 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4321 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4381 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4441 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4501 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4561 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4621 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4681 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4741 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4801 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4861 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
