LOCUS       sequence-470876-        6050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2193m_3xC2 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 6050)
  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 6050)
  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: AiP14563 -
            pAAV-AiE2193m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..6050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             complement(406..417)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(1014..1025)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(1622..1633)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2062..2071
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2068..2070,2071..2073,2074..2784)
                     /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(2113..2134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2374..2393)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2721..2742
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2877..2897)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3084..3195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3313..3453
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3313..3442
                     /label=AAV2 ITR
     rep_origin      3528..3983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3615..3634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3825..3846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4000..4019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4119..4141
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4179..4197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4265..4369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4370..4438,4439..5230)
                     /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(4588..4607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5401..5989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5890..5909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttcaagt ctctgcaagg caaggctagg caacctagct
      181 attaggtcat atcccaggca agtgcttttg gatagccctt gttccagttg ttcaggaccc
      241 acatgaagat caagctaaac atctgctaca tatgagccgg gagttctagg tctagccctg
      301 tatgttcttt gattgatggt tcagactggg agcgccctga gagtccaggt taatggactc
      361 acttgatctt cctgtggcgt ccctatccct ttcggggtcc acaatccttc cttctattgt
      421 tccataagag accccaagtt ccatccactg tttggctgtg ggtgtctgta tatctctgag
      481 tcagctgctg tgtgaagcct ctcagaggaa aatatgctac tttcagtaag catgacagag
      541 tatcattagt agtgctaggg attagtgctt gcacatggga tgggtttcaa gttggaacag
      601 ttattggttg accattccct cagtctctgt tcctctccat gtgcctgtct tacttgtaga
      661 caggattttg agttgaaagt tttgtgggtg ggttggcatc tctatcactc cactggggtt
      721 cctgtctgtc aacaggaggt agcctataca gctcaagtct ctgcaaggca aggctaggca
      781 acctagctat taggtcatat cccaggcaag tgcttttgga tagcccttgt tccagttgtt
      841 caggacccac atgaagatca agctaaacat ctgctacata tgagccggga gttctaggtc
      901 tagccctgta tgttctttga ttgatggttc agactgggag cgccctgaga gtccaggtta
      961 atggactcac ttgatcttcc tgtggcgtcc ctatcccttt cggggtccac aatccttcct
     1021 tctattgttc cataagagac cccaagttcc atccactgtt tggctgtggg tgtctgtata
     1081 tctctgagtc agctgctgtg tgaagcctct cagaggaaaa tatgctactt tcagtaagca
     1141 tgacagagta tcattagtag tgctagggat tagtgcttgc acatgggatg ggtttcaagt
     1201 tggaacagtt attggttgac cattccctca gtctctgttc ctctccatgt gcctgtctta
     1261 cttgtagaca ggattttgag ttgaaagttt tgtgggtggg ttggcatctc tatcactcca
     1321 ctggggttcc tgtctgtcaa caggaggtag cctatacagc tcaagtctct gcaaggcaag
     1381 gctaggcaac ctagctatta ggtcatatcc caggcaagtg cttttggata gcccttgttc
     1441 cagttgttca ggacccacat gaagatcaag ctaaacatct gctacatatg agccgggagt
     1501 tctaggtcta gccctgtatg ttctttgatt gatggttcag actgggagcg ccctgagagt
     1561 ccaggttaat ggactcactt gatcttcctg tggcgtccct atccctttcg gggtccacaa
     1621 tccttccttc tattgttcca taagagaccc caagttccat ccactgtttg gctgtgggtg
     1681 tctgtatatc tctgagtcag ctgctgtgtg aagcctctca gaggaaaata tgctactttc
     1741 agtaagcatg acagagtatc attagtagtg ctagggatta gtgcttgcac atgggatggg
     1801 tttcaagttg gaacagttat tggttgacca ttccctcagt ctctgttcct ctccatgtgc
     1861 ctgtcttact tgtagacagg attttgagtt gaaagttttg tgggtgggtt ggcatctcta
     1921 tcactccact ggggttcctg tctgtcaaca ggaggtagcc tatacagcga gctcgggctg
     1981 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
     2041 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     2101 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2161 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     2221 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     2281 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2341 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2401 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2461 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2521 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     2581 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2641 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     2701 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2761 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     2821 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2881 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2941 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     3001 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3061 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3121 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3181 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3241 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     3301 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3361 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3421 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3481 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3541 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3601 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3661 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3721 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3781 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3841 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3901 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3961 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4021 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4081 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4141 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4201 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4321 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4381 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4441 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4501 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4561 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4621 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4681 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4741 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4801 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4861 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4921 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4981 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5041 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5101 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5161 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5221 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5281 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5341 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5401 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5461 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5521 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5581 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5641 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5701 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5761 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5821 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5881 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5941 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6001 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
