LOCUS       sequence-470744-        5522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1036m 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 5522)
  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 5522)
  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: AiP13877 - pAAV-AiE1036m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..5522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(218..1009,1010..1078))
                     /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     841..860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1079..1183)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1251..1269
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1307..1329)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1429..1448
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1465..1920)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1602..1623)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1814..1833
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1995..2135
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2006..2135
                     /label=AAV2 ITR
     polyA_signal    2253..2364
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2551..2571
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2658..3035)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3066..3131)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3144..3200)
                     /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             complement(join(3210..3920,3921..3923,3924..3926))
                     /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(3252..3273)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3601..3620
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3923..3932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4790..4919
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4981..5522,1..47))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5061..5080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
       61 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      121 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
      181 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
      241 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      301 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
      361 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
      421 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
      481 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      541 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      601 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      661 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      721 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      781 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
      841 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      901 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      961 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1021 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     1081 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     1141 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     1201 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     1261 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1321 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1381 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     1441 agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt
     1501 taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt
     1561 ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc
     1621 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1681 gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1741 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1801 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag
     1861 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     1921 actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     1981 ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     2041 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     2101 agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga
     2161 tctcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
     2221 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
     2281 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
     2341 atggctggca actagaaggc acagtcgaag atctctcgag ccacggaatt gtcagtgccc
     2401 aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca
     2461 agaactaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     2521 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     2581 ataccagtca atctttcaca aattttgtaa tccagaggtt gattatgata tcgaattcgc
     2641 ggccgcggcg cgcctcatta cttagcgctg gtgtacttgg tgatggcctt agtaccctcg
     2701 gacacggcgt gcttggccaa ctccccaggc agcagcaggc gcacggccgt ctggatctcc
     2761 ctggaggtga tggtcgagcg cttgttgtaa tgcgccaggc gggaagcctc acctgcgatg
     2821 cgctcgaaaa tgtcgttcac aaacgagttc atgatgccca tggccttgga cgaaatgccg
     2881 gtgtcagggt ggacctgctt cagaacctta tacacataga tggaatagct ctccttgcgg
     2941 ctgcgcttgc gcttcttgcc gcctttcttc tgcgccttag tcaccgcctt cttggagccc
     3001 tttttcgggg cgggagcaga cttcgctggc tctgggcctc cggagcctcc agatctgagt
     3061 ccggacttgt catcgtcatc cttgtaatcg atgtcatgat ctttataatc tccgtcatgg
     3121 tctttgtagt ctccactagc accaggccct ggattctcct ctacatcccc tgcctgtttc
     3181 aacagtgaga agttggtagc accagagccc ttgtacagct cgtccatgcc gagagtgatc
     3241 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     3301 ctcagcttgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     3361 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg
     3421 cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     3481 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     3541 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     3601 gtcttgtagt tgccgtcgtc tttgaagaag atggtgcgct cctggacgta gccttcgggc
     3661 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     3721 acgccgtagc ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     3781 cagatcagct tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     3841 ctgaacttgt ggccgttgac atcgccgtcc agctcgacca ggatgggcac caccccggtg
     3901 aacagctcct cgcccttgga gaccatggtg gcgaccggta gcgctagctt cgaaagatct
     3961 ggatcccaga agcaaatgta agcaatagat ggctctgccc tgacttttat gcccagcccg
     4021 agctcgtata tctagctaca tttaaggagt agttaattat aaatatgaaa agctacagag
     4081 atacaaaaag atgcatctaa atacactagt aacaagggac tggctttcat gaaaaaacta
     4141 aaagctggtt atatttaagg gaaattgtct tttataagaa tcccacagtg tgagtcaatg
     4201 tgaacgatga aatatgttca atcaactcaa tgtactaatt tgaattggaa agttaatttc
     4261 actaacagca tgtagggcat gaacatgctg ggtcaaaagt cattatagac ttctgaaaga
     4321 cattgttgta cctttctgat catcataatt tattaatttt atagaatttt ggcttatata
     4381 tactttatat tcctgagaat tttttatgga atcataagag aaagaaattt agttataact
     4441 atgatttcac aggcacagga aagaaatata tgattataca tgcataaaca ttgtgctcca
     4501 atatcattag catgcatgga atatgcctta tataaaagca gaggtaggtg gagaaaaaca
     4561 aatatcacag tgtctctcat atccataatt taagtcaaga aaggttctca gtgtaagagt
     4621 tggcaagatt agcagacgat agtaatacag tgaatatgaa tgaagtatga tgtgtgtgtg
     4681 tgtgtatgtt tgtgtgtgat tgtgaatgtt atgatggaag acattatctt atatgctaaa
     4741 taaaaattgg tattttctga attcctctta ggaacacgcg tgcggccgca ggaaccccta
     4801 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4861 aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga
     4921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5401 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5521 tt
//
