LOCUS       sequence-470682-        5596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1429m 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 5596)
  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 5596)
  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: AiP15053 -
            pAAV-AiE1429m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   10..150
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   10..139
                     /label=AAV2 ITR
     misc_feature    157..184
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    185..212
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      281..736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(368..387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     578..599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(753..772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     872..894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(932..950)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1018..1122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1123..1191,1192..1983)
                     /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(1341..1360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2154..2742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2643..2662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2799..2826
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2827..2854
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2861..2990
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3809..3818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3815..3817,3818..3820,3821..4531)
                     /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(4121..4140)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4468..4489
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4541..4597
                     /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             4610..4675
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4706..5083
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(5170..5190)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5377..5488
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
       61 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
      121 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctatc agcacacaat tgcccattat
      181 acgcgcgtat aatggactat tgtgtgctga tatgatgcgg tattttctcc ttacgcatct
      241 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
      301 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
      361 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      421 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      481 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      541 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      601 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
      661 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      721 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      781 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
      841 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
      901 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
      961 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     1021 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1081 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1141 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1201 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1261 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1321 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1381 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1441 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1501 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1561 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1621 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1681 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1741 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1801 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1861 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1921 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1981 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2041 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2101 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2161 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2221 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2281 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2341 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2401 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2461 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2521 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2761 tttttacggt tcctggcctt ttgctggcct tttgctcata tcagcacaca attgcccatt
     2821 atacgcgcgt ataatggact attgtgtgct gataacatgt cctgcaggca gctgcgcgct
     2881 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     2941 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac
     3001 gcgtcagcgg ttgggtggga atgattctgc cctggttgtt tctctatttc ctttgctcca
     3061 tccttccctc tgcatttcaa atatgcatat tacagatctt cagtcccttc tttccctcat
     3121 gtatatgaac ttctgggtga cctcaactta atttgaatat cattgccata ccaatgaata
     3181 ctatacgatg acattgctcc acattacaca ctctctgctt ttggcctaat gtgcctaggg
     3241 tgctatttat actgatgtac ctctacattc attcatctgt agagatgcat ctctacagaa
     3301 tacacattga ttctcatcta aatttctaca gcctgaatga aaactcatgt tttcctctcc
     3361 cttgacttct ctccaaaact caataacaaa atatcttttt gcttatgtct cttaatttcc
     3421 tcctttcaca agcagccctg tgactgaata ctttaattta tccacatgcc taaacacatt
     3481 ttctgtaatt actcccagtg cttctcattg cttattttat tctctgagtc tctttgagat
     3541 gaactgtgca taacagatga tataattttg tatggagtaa agatagttat gaaatatgaa
     3601 acgaagagtc ataaaagtat ctttgcagca gcaaaaagaa tatctatata tgtacatata
     3661 ttttgagtgt gtatagatat ataaatatct atgtgcatat acatattcat gttctgagct
     3721 cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc
     3781 agatctttcg aagctagcgc taccggtcgc caccatggtc tccaagggcg aggagctgtt
     3841 caccggggtg gtgcccatcc tggtcgagct ggacggcgat gtcaacggcc acaagttcag
     3901 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     3961 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt
     4021 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4081 gcccgaaggc tacgtccagg agcgcaccat cttcttcaaa gacgacggca actacaagac
     4141 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4201 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4261 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     4321 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     4381 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
     4441 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4501 gatcactctc ggcatggacg agctgtacaa gggctctggt gctaccaact tctcactgtt
     4561 gaaacaggca ggggatgtag aggagaatcc agggcctggt gctagtggag actacaaaga
     4621 ccatgacgga gattataaag atcatgacat cgattacaag gatgacgatg acaagtccgg
     4681 actcagatct ggaggctccg gaggcccaga gccagcgaag tctgctcccg ccccgaaaaa
     4741 gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc gcaagcgcag
     4801 ccgcaaggag agctattcca tctatgtgta taaggttctg aagcaggtcc accctgacac
     4861 cggcatttcg tccaaggcca tgggcatcat gaactcgttt gtgaacgaca ttttcgagcg
     4921 catcgcaggt gaggcttccc gcctggcgca ttacaacaag cgctcgacca tcacctccag
     4981 ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc acgccgtgtc
     5041 cgagggtact aaggccatca ccaagtacac cagcgctaag taatgaggcg cgccgcggcc
     5101 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5161 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5221 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     5281 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5341 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     5401 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5461 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5521 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     5581 tctcacgtgc ggaccg
//
