LOCUS       sequence-466235-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2067m 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 4731)
  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 4731)
  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: AiP1282 - pAAV-AiE2067m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   269..398
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1011..1020
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1017..1019,1020..1022,1023..1733)
                     /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(1062..1083)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1323..1342)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1670..1691
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1826..1846)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2033..2144
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2262..2402
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2262..2391
                     /label=AAV2 ITR
     rep_origin      2477..2932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2564..2583)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2774..2795
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2949..2968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3068..3090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3128..3146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3214..3318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3319..3387,3388..4179)
                     /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(3537..3556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4350..4731,1..207)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
       61 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      241 gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact
      301 gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
      361 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc gtcagagagt
      421 tcagttgcaa tcaactgtgt ttaatattca cacattgtaa tttaaacatt aaagatctca
      481 agcaaagatc acgatcttag aaggtattaa attcaaaaag cccactcccc cggaattgca
      541 cagttagctc ctgatttctg gtttatgtcc cattagccca tggtgataag tatggactca
      601 tataaaggaa aagcacgaat ccccatttca gaacattaag tctagatttt tgacgataac
      661 ctatttttgt gctgctatta gggggcacta gcctgtaccc acatccagtt attaatctgt
      721 gcaacattgt ctctctggcg tgttctgttc tttctaagtc tgctgaactt aagtgtcacc
      781 aaataagtgg gttattaaac gggatcgaga aatgaatcaa tcaagtatat tcaaatgctc
      841 agtctccctc attgttcttt gacttcatta tttaggcatt aaattccatg caacactgga
      901 ggtaattgtc aatacaggag ctcgggctgg gcataaaagt cagggcagag ccatctattg
      961 cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc gccaccatgg
     1021 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1081 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1141 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1201 tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1261 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1321 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1381 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1441 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     1501 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     1561 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1621 tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1681 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg
     1741 acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg
     1801 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1861 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1921 aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1981 caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct
     2041 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2101 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2161 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2221 aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     2281 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2341 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2401 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     2461 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2521 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     2581 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2641 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2701 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2761 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     2821 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     2881 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     2941 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3001 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3061 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3121 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3181 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3241 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3301 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3361 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3421 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3481 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3541 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3601 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3661 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3721 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3781 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3841 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3901 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3961 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4021 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4081 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4141 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4201 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4261 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4321 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4381 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4441 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4501 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4561 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4621 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4681 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct a
//
