LOCUS       sequence-466272-        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2270m 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 4835)
  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 4835)
  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: AiP1603 - pAAV-AiE2270m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     88..107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(124..579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(261..282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     473..492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   654..794
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   665..794
                     /label=AAV2 ITR
     polyA_signal    912..1023
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1210..1230
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1323..2033,2034..2036,2037..2039))
                     /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(1365..1386)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1714..1733
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1973..1994
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2036..2045
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2762..2891
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2953..3541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3033..3052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3712..4503,4504..4572))
                     /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     4335..4354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4573..4677)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4745..4763
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4801..4823)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
       61 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
      121 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
      181 cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
      241 agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
      301 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
      361 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
      421 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
      481 aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
      541 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
      601 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag
      661 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
      721 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      781 cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg ctattgtctt
      841 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
      901 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
      961 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
     1021 cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc ccctgtccag
     1081 cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca ggatttatac
     1141 aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca
     1201 gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa
     1261 attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc gccgtcgact
     1321 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     1381 accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta gctcaggtag
     1441 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     1501 gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc cttgatgccg
     1561 ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     1621 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     1681 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     1741 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     1801 ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg
     1861 gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt cagcttgccg
     1921 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     1981 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     2041 gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat gtaagcaata
     2101 gatggctctg ccctgacttt tatgcccagc ccgagctccg ccggcgtgga ggccggcccg
     2161 ctcccccaga ggtcctggcc tcaatgcctg ggacggagga tgctgtccgc cgccgcactg
     2221 ccccgctggt tctgggggcg ggtgaagctg ggtcctgcca ggcacagctg gagaggaagg
     2281 gccgggcggg aggggggaag aggagggctg agaagagaag gaggggtggt gaggcttccc
     2341 agcttcgagc tggggaggca ggcttgagct gcgctgcagg agcctggacc agtccacccg
     2401 ccacccgcca ccccgcacag agggactctg gcattctcgc cccgggggcc cgcgctgcgc
     2461 ctctctcttc cttatccgtc cgccctctca atcccgccaa gtgctgcgcc aaggccaagt
     2521 ctgctctcct cctgaacttt cccaggactc taacctacgg ttagattcca ccgcggggct
     2581 cccggacacc cgctgcggaa gctgggttgg ggagagggcg tgaccacaag agctcagggg
     2641 ccccgagcca gcagtgctga acccgggctg gacagagctt gctttcctgc gtccccagaa
     2701 catgagtgct ggggagggct tctgcgccgg cgattattac tcttacaacg cgtgcggccg
     2761 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2821 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2881 ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3361 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4681 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4741 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4801 cctctgacac atgcagctcc cggagacggt cacag
//
