LOCUS       sequence-473478-        5862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1431m 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 5862)
  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 5862)
  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: AiP15055 -
            pAAV-AiE1431m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5862
                     /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      1215..1224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1221..1223,1224..1226,1227..1937)
                     /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(1527..1546)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1874..1895
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1947..2003
                     /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             2016..2081
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2112..2489
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2576..2596)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2783..2894
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3012..3152
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3012..3141
                     /label=AAV2 ITR
     misc_feature    3159..3186
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3187..3214
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3283..3738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3370..3389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3580..3601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3755..3774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3874..3896
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3934..3952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4020..4124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4125..4193,4194..4985)
                     /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(4343..4362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5156..5744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5645..5664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5801..5828
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5829..5856
                     /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 gcgtgacaat gctatctcaa tgtataagat gactactggc
      181 ctatatttac ctgacaagaa tctattcctc cttcttcata tcctgcacat atcatacttt
      241 cagtgatgtt gtattctggc agctgttgct ggcacttttc attggatata agaggaacat
      301 cagcttcttt cagcacatca acagtagaac ctgcttgaaa tgggaaatgc agacagccag
      361 tcagtaatga ttaacaggca tcctcgaatg aaacgccatc catctgattt atcccagcat
      421 tttgctctca ttatcaagta ctatgtggat gtataatgga actaactacc aaccttgcac
      481 tttcaagtac ataataaacc catctgtgca ctttggtaaa tggagagcat ttctttcttc
      541 ctttcctaag gggtagtaag aattgtggat tggactaggg tggtctttta ggcattctca
      601 gtatttttta aagagttaca gtgttgctgg tagaaaggaa gtctgccagc acaaagtgtt
      661 cattcatgga aatcagcaca gatttggata tccagaagag cttttcatcc agcacctgag
      721 gaggtgctcc ctcctgtata aaggcattca gctcctttgt ggttctgtgt gtgcgagagc
      781 aggtgcacgc tgaacagggg acaaagccag ctgaactgtg ctctactctt ggggcaatat
      841 gcttgccatc ccaggcaggt gaagagaaag gaagcatatt tcagcattgt gtgtttgcat
      901 ttgttttatc acaaatcact cactgcaagg atgaacctct ccttactaaa ctactgttcc
      961 caagaagctt gtgaagcaag caggtgttag taccttcaga aaaggaacat aagcatttat
     1021 tcatgattag caattctctt tgatgtcaca aaagggatag aaacttaagt gtaacttatt
     1081 atatctgtca ggcagtattg taaaactata tacaatccca tgagctcggg ctgggcataa
     1141 aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc
     1201 tagcgctacc ggtcgccacc atggtctcca agggcgagga gctgttcacc ggggtggtgc
     1261 ccatcctggt cgagctggac ggcgatgtca acggccacaa gttcagcgtg tccggcgagg
     1321 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     1381 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
     1441 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1501 tccaggagcg caccatcttc ttcaaagacg acggcaacta caagacccgc gccgaggtga
     1561 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1621 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1681 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     1741 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1801 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     1861 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1921 tggacgagct gtacaagggc tctggtgcta ccaacttctc actgttgaaa caggcagggg
     1981 atgtagagga gaatccaggg cctggtgcta gtggagacta caaagaccat gacggagatt
     2041 ataaagatca tgacatcgat tacaaggatg acgatgacaa gtccggactc agatctggag
     2101 gctccggagg cccagagcca gcgaagtctg ctcccgcccc gaaaaagggc tccaagaagg
     2161 cggtgactaa ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc aaggagagct
     2221 attccatcta tgtgtataag gttctgaagc aggtccaccc tgacaccggc atttcgtcca
     2281 aggccatggg catcatgaac tcgtttgtga acgacatttt cgagcgcatc gcaggtgagg
     2341 cttcccgcct ggcgcattac aacaagcgct cgaccatcac ctccagggag atccagacgg
     2401 ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag ggtactaagg
     2461 ccatcaccaa gtacaccagc gctaagtaat gaggcgcgcc gcggccgcga attcgatatc
     2521 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2581 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2641 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     2701 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2761 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2821 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2881 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2941 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     3001 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3061 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3121 agtgagcgag cgagcgcgca gctgcctgca ggggcgccta tcagcacaca attgcccatt
     3181 atacgcgcgt ataatggact attgtgtgct gatatgatgc ggtattttct ccttacgcat
     3241 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     3301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     3361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3481 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3601 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     3661 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3721 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3781 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3841 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3901 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3961 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4021 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4081 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4141 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4201 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4261 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4321 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4381 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4441 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4501 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4561 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4621 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4681 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4741 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4801 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4861 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4921 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4981 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5041 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5101 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5161 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5221 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5281 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5341 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5401 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5461 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5521 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5581 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5641 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5701 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5761 cctttttacg gttcctggcc ttttgctggc cttttgctca tatcagcaca caattgccca
     5821 ttatacgcgc gtataatgga ctattgtgtg ctgataacat gt
//
