LOCUS       sequence-470472-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0765m 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 4682)
  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 4682)
  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: AiP12600 - pAAV-AiE0765m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(237..341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     409..427
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(465..487)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     587..606
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(623..1078)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(760..781)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     972..991
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1153..1293
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1164..1293
                     /label=AAV2 ITR
     polyA_signal    1411..1522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1709..1729
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1822..2532,2533..2535,2536..2538))
                     /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(1864..1885)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2213..2232
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2472..2493
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2535..2544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3108..3237
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3299..3887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3379..3398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4058..4682,1..167,168..236))
                     /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     join(4681..4682,1..18)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
       61 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
      121 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
      181 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
      241 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      301 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      361 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
      421 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
      481 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
      541 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
      601 attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta
      661 aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat
      721 aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca
      781 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
      841 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
      901 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
      961 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg
     1021 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac
     1081 tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     1141 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     1201 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     1261 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
     1321 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     1381 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     1441 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     1501 ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa
     1561 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag
     1621 aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     1681 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     1741 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg
     1801 ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     1861 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     1921 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1981 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
     2041 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
     2101 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     2161 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     2221 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     2281 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     2341 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     2401 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     2461 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     2521 ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca
     2581 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctcgcg
     2641 aggctaatgg tgcgtaaaag cactggtgag atctgggggc gcctccttgg ctgacgtcag
     2701 agagagagtt taaaaagggg agaccgtgga gagctccata gcggctgaag gagacgctac
     2761 cgaagccgtc gctgctgcct gaggacctgc gactagactg acccaccgcg ctccagcttg
     2821 gctgcctgag gcaaggaaga tgctgtcctg ccgtctccag tgcgccctgg ctgcgctctg
     2881 catcgtcctg gctttgggcg gtgtcaccgg cgcgccctcg gaccccagac tccgtcagtt
     2941 tctgcagaag tctctggcgg ctgccaccgg gaaacaggta aggactgtcc cccttacgaa
     3001 tcccccagcc ttcctgctgc agcccctgcg acaggtgttt agcgggcgct tctcagagtc
     3061 gctcagcccc taagctccca gggaaacttt tgaacgcgtg cggccgcagg aacccctagt
     3121 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3181 ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca
     3241 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3721 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3961 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4681 at
//
