LOCUS       sequence-470747-        5190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1035m 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 5190)
  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 5190)
  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: AiP14024 -
            pAAV-AiE1035m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      78..533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(165..184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     375..396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(550..569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     669..691
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(729..747)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        815..919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(920..988,989..1780)
                     /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(1138..1157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1951..2539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2440..2459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2601..2730
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3256..3265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3262..3264,3265..3267,3268..3978)
                     /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(3568..3587)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3915..3936
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3988..4044
                     /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             4057..4122
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4153..4530
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4617..4637)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4824..4935
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   join(5053..5190,1..3)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5053..5182
                     /label=AAV2 ITR
ORIGIN
        1 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
       61 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      121 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
      181 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      241 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
      301 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      361 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
      421 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
      481 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
      541 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
      601 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
      661 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
      721 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
      781 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      901 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      961 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1021 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1081 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1141 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1201 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1261 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1321 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1381 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1441 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1501 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1561 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1621 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1681 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1741 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1801 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1861 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1921 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1981 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2041 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2101 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2161 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2221 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2281 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2341 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2401 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2461 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2521 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2581 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     2641 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2701 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtcaccta cttatattat
     2761 gtatgcttta aaatgtagtc atatttattg tgatataatt tatgttattc tctgtccatg
     2821 gttcctgtct cattcttcct gaaaccctca taattttcta ttagcaatat taacacacat
     2881 tttataataa tacttctgtt tccagtgtac agttcattca atcatctatc gatgggcacc
     2941 tacagtgagt ctgtatccta tctaatgtaa atagagccat aatgaatata ggattgttgg
     3001 catctctttt ataagttata tccatcttca tcataaatat gcatggggct ggtaggcttg
     3061 gatcatagtg taattccatt tatagtttaa ttcattttta tacaagtctt tttccggggg
     3121 cacaggcttg cctgaaactt atgatactcc atgcccttac tggagctcgg gctgggcata
     3181 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
     3241 ctagcgctac cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg
     3301 cccatcctgg tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag
     3361 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     3421 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     3481 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3541 gtccaggagc gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg
     3601 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3661 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3721 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     3781 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3841 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     3901 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3961 atggacgagc tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg
     4021 gatgtagagg agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat
     4081 tataaagatc atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga
     4141 ggctccggag gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag
     4201 gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc
     4261 tattccatct atgtgtacaa ggttctgaag caggtccacc ctgacaccgg catttcgtcc
     4321 aaggccatgg gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat cgcaggagag
     4381 gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctcccggga gatccagacg
     4441 gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag
     4501 gccatcacca agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat
     4561 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4621 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4681 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     4741 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4801 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4861 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4921 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4981 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     5041 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5101 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5161 cagtgagcga gcgagcgcgc agctgcctgc
//
