LOCUS       sequence-470777-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0365h 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 5623)
  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 5623)
  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: AiP14524 -
            pAAV-AiE0365h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    171..198
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    199..226
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     rep_origin      complement(283..871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(363..382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1042..1833,1834..1902))
                     /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     1665..1684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1903..2007)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2075..2093
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2131..2153)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2253..2272
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2289..2744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2426..2447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2638..2657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     misc_feature    2813..2840
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2841..2868
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2875..3015
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2886..3015
                     /label=AAV2 ITR
     polyA_signal    3133..3244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3431..3451
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3538..3915)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3946..4011)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4024..4080)
                     /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(4090..4800,4801..4803,4804..4806))
                     /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(4132..4153)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4481..4500
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      4803..4812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctttgctcat tccaaatgca acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc
       61 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
      121 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt tatcagcaca
      181 caatagtcca ttatacgcgc gtataatggg caattgtgtg ctgatatgag caaaaggcca
      241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2041 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2101 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2161 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2221 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     2281 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2341 cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata
     2401 gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2461 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2521 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2581 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2641 gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt
     2701 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     2761 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat catatcagca
     2821 cacaatagtc cattatacgc gcgtataatg ggcaattgtg tgctgatagg cgcccctgca
     2881 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     2941 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     3001 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     3061 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     3121 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     3181 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     3241 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     3301 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     3361 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     3421 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     3481 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgcctcatta
     3541 cttagcgctg gtgtacttgg tgatggcctt agtaccctcg gacacggcgt gcttggccaa
     3601 ctccccaggc agcagcaggc gcacggccgt ctggatctcc ctggaggtga tggtcgagcg
     3661 cttgttgtaa tgcgccaggc gggaagcctc acctgcgatg cgctcgaaaa tgtcgttcac
     3721 aaacgagttc atgatgccca tggccttgga cgaaatgccg gtgtcagggt ggacctgctt
     3781 cagaacctta tacacataga tggaatagct ctccttgcgg ctgcgcttgc gcttcttgcc
     3841 gcctttcttc tgcgccttag tcaccgcctt cttggagccc tttttcgggg cgggagcaga
     3901 cttcgctggc tctgggcctc cggagcctcc agatctgagt ccggacttgt catcgtcatc
     3961 cttgtaatcg atgtcatgat ctttataatc tccgtcatgg tctttgtagt ctccactagc
     4021 accaggccct ggattctcct ctacatcccc tgcctgtttc aacagtgaga agttggtagc
     4081 accagagccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     4141 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actggtagct
     4201 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     4261 gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttggcctt
     4321 gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt agttgtactc
     4381 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     4441 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     4501 tttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     4561 gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc ccagggtggt
     4621 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct tcagggtcag
     4681 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgttgac
     4741 atcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgga
     4801 gaccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga agcaaatgta
     4861 agcaatagat ggctctgccc tgacttttat gcccagcccg agctcctttc cttttttatt
     4921 ttaacaagtg tttctgtgta ggaaacacag gctgaaacct gctttgggta acatgctctg
     4981 aatttagatg accacaaaat ctattttgag aaataatcgc tcacttatta accactcact
     5041 cgtgccaaca cctagtaggt gggcgctcaa taagtgtttg ttggccaaag gaatgactat
     5101 tggggacgga ccccagcctc actggcaggg agagcctggc ccccagcacc ttgccgggga
     5161 ggctcctctg ggcctgactg agccccattc agcccccatg tgcagaggat gactcagcct
     5221 gtgacttcat tcacccagcc tggtgtttat ccactcacca caaaccggtt gatactgttt
     5281 ggagctaccc cgaggtgacg ctctgctggt ggagaatgac atgcagtcag cttgtggcct
     5341 ctggggccaa aacagctgaa atgctgtggc ttctgaagaa cccattgtct gctagtcaag
     5401 cgtgaggaca aggaaggttc tggtgctgcc ccccagctcc ctctgagagg cattttggct
     5461 tccatgtggg tgcgggacag accacagcag ggggccctag cagggctctg accccaagcg
     5521 tggaagaatg gaccctggca tggggatccc tcatagaagc ccctcaaaaa atactgtggg
     5581 ggtccagata gagctggaag gagctgagga gtttagaagg cag
//
