LOCUS       sequence-466242-        5083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2081m 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 5083)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5083)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP1359 - pAAV-AiE2081m-minBG-SYFP2-4X2C-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      273..861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     762..781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   923..1052
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1797..1806
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1803..1805,1806..1808,1809..2519)
                     /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(1848..1869)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2109..2128)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2456..2477
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2832..2852)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3039..3150
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3268..3408
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3268..3397
                     /label=AAV2 ITR
     rep_origin      3483..3938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3570..3589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3780..3801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3955..3974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4074..4096
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4134..4152)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4220..4324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4325..4393,4394..5083,1..102)
                     /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(4543..4562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
       61 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
      121 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
      181 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
      241 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
      301 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
      361 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
      421 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
      481 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      541 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      601 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      661 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      721 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      781 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      841 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      901 tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc
      961 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     1021 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtatac tgtaaaaagc
     1081 aagaagaacc aatcacttaa tctaaaattt agaactctga agtgattcat tccactgacc
     1141 attctctttt cccagttctc ttatcttgtc agatacacag gaagctcagg acagttttgc
     1201 tcatcccttt gtttctagca gttctgagta atgaattcaa atagcagtta ttatctcaat
     1261 gccatgacaa ttggctaagc taactaccca gttcacttat cgagggagca aaacactaga
     1321 tggccaataa gaatgacaga cagaatgtat gattgatgtg aaacacccag agaatcaagt
     1381 gctttcaaag agttaaatgt gttctccatt atggctgaca gctggagaaa ctatcattgg
     1441 aaactgagat gaattgtttg ctctgtgcag gtgggagaag agaattaaat gacagacaca
     1501 tgaaactgtg ctttgcatct tcttgtcagc actactctgt tggccaagag tgaaatgcaa
     1561 tttcagttgg cttgaacgcc acaggatttt acatctgaat caactaagtt ctaattatgg
     1621 gcagatttat ctactaatac catcaccata acaaccaaaa ccatgcccca tgcctttcat
     1681 gtcttcacta tttcactgag tttgagctcg ggctgggcat aaaagtcagg gcagagccat
     1741 ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca
     1801 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1861 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1921 acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
     1981 ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga
     2041 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2101 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2161 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2221 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
     2281 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     2341 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2401 actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     2461 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2521 aagtcgacgg cgcgccgcgg ccttaaagag accggttcac tgtgacagta aaagagaccg
     2581 gttcactgtg agaatgaaag agaccggttc actgtgatcg gaaaagagac cggttcactg
     2641 tgagcggcct tgaaacccag cagacaatgt agctcagtag aaacccagca gacaatgtag
     2701 ctgaatggaa acccagcaga caatgtagct tcggagaaac ccagcagaca atgtagctgt
     2761 cgacgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2821 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2881 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     2941 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3001 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     3061 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3121 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3181 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     3241 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3301 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3361 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3421 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3481 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3541 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3661 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     3721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3781 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3841 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3901 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3961 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4021 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4081 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4141 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4321 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4381 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4441 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4501 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4561 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4621 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4681 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4741 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4801 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4861 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4921 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4981 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5041 gtctcgcggt atcattgcag cactggggcc agatggtaag ccc
//
