LOCUS       sequence-461942-        5454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2173m 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 5454)
  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 5454)
  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: AiP1440 - pAAV-AiE2173m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1466..1475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1472..1474,1475..1477,1478..2188)
                     /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(1517..1538)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1778..1797)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2125..2146
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2281..2301)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2488..2599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2717..2857
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2717..2846
                     /label=AAV2 ITR
     rep_origin      2932..3387
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3019..3038)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3229..3250
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3404..3423)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3523..3545
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3583..3601)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3669..3773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3774..3842,3843..4634)
                     /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(3992..4011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4805..5393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5294..5313
                     /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 gcgttgtaag agtaataatc gccggcgcca cttgagaagt
      181 ctggaaagga gataaaatgt gggagaccca gtgtctactg ttggccctaa ttggcttagg
      241 acgatctagc agatattgca gggcctacag gaagagccaa gcctggcagc cttgggacca
      301 aagtcctcca cctgacaaat tcaccctcaa aattagggat ccacgctctc gctattctga
      361 ccccccgaag aggggactct tggccaccag aggggactga gaagaaatcg gtattattag
      421 gctaaaagaa cagtggacta cttcagcaag gcctagcgac cggagcagca gaagccttcc
      481 atggtagtgc tagcctttct ccctcccagc cccagtttcc tctgggcagg ggcctggccc
      541 cctgtcttcc cagggctgcc ctttgagaac cacctgcccc agccggccag agacttcctg
      601 tagcgcaaga gatttatgca aacgggctgg ggcggtgatg tcaccccaag gggactatct
      661 cccagtggca ggcccttcga taaaatcagg aacttgtgct ggccctgcaa tgtcaaggga
      721 gggggctcac ccagggctcc tgtagctcag ggggcaggcc tgagccctgc gtctgaccca
      781 cgaccgtcca gtcccccgac ggggcacctg gtcctgaggg ggatccgcct tgatccccac
      841 cgaagcaggg gatctgacca acctggagct cagctggatg ttacaggcgt gcaaaatgga
      901 agggttttcc ctcaccgccc ctgtgagtac tggggcttca ctaccagccc agcctggggc
      961 tttgggatgt gtgaccatct gcccactaac ccaccagccc acccgatgcc ctccggctcc
     1021 ctatcagtgg gaagttggtg tttcctcccc ggagctgctg gtctgagacc cagcgagggg
     1081 ccattggctt ccttagagca tgccctggcg ggctgagggt tgccggctgg ccccatggtg
     1141 gggctgtgat cggtggggcc tgaacagcta tttatagatg ggttcagtgg caggccgaac
     1201 tctagggcag ggttctgggt tgaagatggc gagggcggct cctgagaggc catggatgag
     1261 gagatctgga gactggccta ggagtctctg gaggcaacag gagaggtttc aactcaagac
     1321 ctgtgggctc ctgaggcaaa ggctattaga actcgaagtc accccgccgg cggagctcgg
     1381 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
     1441 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1501 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1561 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     1621 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
     1681 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1741 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1801 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1861 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1921 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     1981 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2041 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     2101 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2161 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     2221 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2281 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2341 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     2401 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2461 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2521 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2581 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2641 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2701 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2761 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2821 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2881 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2941 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3001 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3061 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3121 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3181 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3241 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3301 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3361 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3421 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3481 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3541 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3601 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3661 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3721 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3781 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3841 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3901 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3961 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4021 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4081 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4141 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4201 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4261 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4321 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4381 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4441 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4501 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4561 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4621 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4681 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4741 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4801 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4861 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4921 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4981 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5041 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5101 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5161 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5221 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5281 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5341 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5401 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
