LOCUS       sequence-473573-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0346m 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 5351)
  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 5351)
  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: AiP14539 -
            pAAV-AiE0346m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5351
                     /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      704..713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(710..712,713..715,716..1426)
                     /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(1016..1035)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1363..1384
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1436..1492
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1505..1570
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1601..1978
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2065..2085)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2272..2383
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2501..2641
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2501..2630
                     /label=AAV2 ITR
     misc_feature    2648..2675
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2676..2703
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2772..3227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2859..2878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3069..3090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3244..3263)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3363..3385
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3423..3441)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3509..3613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3614..3682,3683..4474)
                     /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(3832..3851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4645..5233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5134..5153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5290..5317
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5318..5345
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacacc aagtgtctac ccccagatcc tcatagaaga
      181 aaaactaaac tcttgcttgc acaaagcact gtagcttgac ttttcacttt catctgaaag
      241 catttctgcc tgataaaaga actcagtgtg taaacaagat gtgcaattcc catcttcagt
      301 gaatagcaat cctttgaacg tattccaggg atgactgaga gaatgctaaa tacacctgca
      361 gcctcagatc agattgtcta tgcttccagc ctccaccggt gtacgaaaac tgtgaaataa
      421 gattcctaaa atatagcagt tgccgaactg agtgactggc aaataatgac ctgttgcact
      481 ttctatttaa atatgagatt ttctatcaaa atacatcatc tccattacaa aatatatttt
      541 ccaaaaaaaa aaaaaagtat ttcatcaaag ttgtattcat cccaacagta ttttagccac
      601 ctttctgtgg gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
      661 ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtctccaa
      721 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgatgtcaa
      781 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      841 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      901 cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
      961 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaagacga
     1021 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1081 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1141 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1201 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1261 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1321 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1381 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggct ctggtgctac
     1441 caacttctca ctgttgaaac aggcagggga tgtagaggag aatccagggc ctggtgctag
     1501 tggagactac aaagaccatg acggagatta taaagatcat gacatcgatt acaaggatga
     1561 cgatgacaag tccggactca gatctggagg ctccggaggc ccagagccag cgaagtctgc
     1621 tcccgccccg aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga aaggcggcaa
     1681 gaagcgcaag cgcagccgca aggagagcta ttccatctat gtgtataagg ttctgaagca
     1741 ggtccaccct gacaccggca tttcgtccaa ggccatgggc atcatgaact cgtttgtgaa
     1801 cgacattttc gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca acaagcgctc
     1861 gaccatcacc tccagggaga tccagacggc cgtgcgcctg ctgctgcctg gggagttggc
     1921 caagcacgcc gtgtccgagg gtactaaggc catcaccaag tacaccagcg ctaagtaatg
     1981 aggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga
     2041 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2101 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2161 atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2221 aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt
     2281 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2341 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2401 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2461 atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     2521 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2581 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2641 gggcgcctat cagcacacaa ttgcccatta tacgcgcgta taatggacta ttgtgtgctg
     2701 atatgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2761 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2821 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     2881 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2941 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3001 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3061 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3121 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3181 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3241 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3301 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3361 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5281 ttttgctcat atcagcacac aattgcccat tatacgcgcg tataatggac tattgtgtgc
     5341 tgataacatg t
//
