LOCUS       sequence-432595-        5048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2455m 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 5048)
  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 5048)
  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: AiP1923 - pAAV-AiE2455m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   328..457
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(519..1107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(599..618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1278..2069,2070..2138))
                     /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     1901..1920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2139..2243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2311..2329
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2367..2389)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2489..2508
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2525..2980)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2662..2683)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2874..2893
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3055..3195
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3066..3195
                     /label=AAV2 ITR
     polyA_signal    3313..3424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3611..3631
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3724..4434,4435..4437,4438..4440))
                     /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(3766..3787)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4115..4134
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4374..4395
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4437..4446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gatggatggt caggagtttc actctgtcgg ctcagccctg tgaagcagag actaaggatt
       61 tttcagtgtt agtatctggg catggaaaag gaatacagtt tgaataaaaa gcacagtgta
      121 ctggttagat tttcatcaaa ttgacacaaa ctaaaatact ctggggaaag ggaggatcaa
      181 cagagaaaat gtctccatca gactggcatg tatgcaaatc tgtggagcat tttcttgact
      241 aatgattgat atgggagggc ccagcataat gtggtcagtg acagccatag ctggtgtgta
      301 taagaaagca tacacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct
      361 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
      421 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa
      481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1561 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1861 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2281 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     2341 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2401 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2461 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2521 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2581 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     2641 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2701 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2761 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2821 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2881 aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2941 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     3001 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
     3061 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     3121 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     3181 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     3241 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     3301 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     3361 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     3421 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     3481 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     3541 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     3601 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     3661 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
     3721 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     3781 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
     3841 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     3901 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
     3961 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     4021 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     4081 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     4141 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     4201 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     4261 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     4321 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     4381 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     4441 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     4501 agatggctct gccctgactt ttatgcccag cccgagctcc gccggcgggc tccagtgttc
     4561 agcttctgaa gcatgtttgt ctacagaaag cagctctttc tgtctgcttt gcactgtgat
     4621 ttgaaatgtc gtcaacgctt tgatgcagct gcctctgttc agacctctaa gaaacaaagc
     4681 aaatgtttat tcattctcaa ttggtattgt ttgaatctga aatgttccca aagtcctgcg
     4741 tattgaagac tcacttacca gtctgtatgc tgttgaaaaa tgttggatcc tttaagaggc
     4801 agggctcgct aggatggggt tagggacttg ggtgtatctc cccaaaacag tgcatagtgc
     4861 ccccttctct ccttgtttcc tctacaatat gaagtgaaca ttaacaagcc ccttctgcta
     4921 tttgcttcca ggaccaagaa atcatggctt gaaagcttta agcaatcctt tcctcttttg
     4981 aagttgactc agcaacagaa aagccactag cactcagcct tgctacttgc tctgaaaact
     5041 gcctcctg
//
