LOCUS       sequence-470772-        5596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0358h 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: AiP14522 -
            pAAV-AiE0358h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      122..710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     611..630
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    767..794
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    795..822
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   829..958
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1777..1786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1783..1785,1786..1788,1789..2499)
                     /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(2089..2108)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2436..2457
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2509..2565
                     /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             2578..2643
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2674..3051
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3138..3158)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3345..3456
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3574..3714
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3574..3703
                     /label=AAV2 ITR
     misc_feature    3721..3748
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3749..3776
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3845..4300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3932..3951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4142..4163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4317..4336)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4436..4458
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4496..4514)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4582..4686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4687..4755,4756..5547)
                     /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(4905..4924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
       61 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
      121 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
      181 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      241 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      601 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      661 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      721 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcatatc agcacacaat
      781 tgcccattat acgcgcgtat aatggactat tgtgtgctga taacatgtcc tgcaggcagc
      841 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
      901 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
      961 ggccgcacgc gtgattaatg acactctaat ggtacagaac agaaaattct ctgtctctaa
     1021 gatcatctaa accaaagaca tgtacacaac tacctataat gtaggccaga ccacagaaat
     1081 gctaagtgat gctttaaagt cttctgaaaa ataatgcctt aactaaacaa agtcccccta
     1141 catccagagt atgaatgatg ctgctattac ttatttctac tgtcttccat ctattttata
     1201 gtgttctttg atccattctt tgcattcttc ttgtttgaaa gtaatattct tgatagttta
     1261 ataatgagac ccctcctttg ttcctagatg ccttggcggc taatcaacaa aagccattga
     1321 aagacttaat cctctaagct tggcaccaca tcagtctgga agtagtttac atgcagagtc
     1381 ctttggagta aatactcatt agaggttctg ttggctcatt tggtcaccca cattttgcag
     1441 tgggtggcct agttggcatc ctcattattt tctttttcca ctcctgaaaa taatccagat
     1501 gtagcaacct gacactactt tctttggtac tccaaatcca ttgttatgaa gggtccatta
     1561 atttgtgatg ctagctaata tgtccaaata aataaaggcc tcaggccagt aatgctgtaa
     1621 taaatctttt actaatacac ttttttcagt gcaacctatt gaatgcaatt caatggcttc
     1681 aaggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
     1741 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtctc caagggcgag
     1801 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgatgt caacggccac
     1861 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1921 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     1981 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2041 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaaaga cgacggcaac
     2101 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2161 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2221 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     2281 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     2341 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     2401 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2461 gccgccggga tcactctcgg catggacgag ctgtacaagg gctctggtgc taccaacttc
     2521 tcactgttga aacaggcagg ggatgtagag gagaatccag ggcctggtgc tagtggagac
     2581 tacaaagacc atgacggaga ttataaagat catgacatcg attacaagga tgacgatgac
     2641 aagtccggac tcagatctgg aggctccgga ggcccagagc cagcgaagtc tgctcccgcc
     2701 ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc
     2761 aagcgcagcc gcaaggagag ctattccatc tatgtgtata aggttctgaa gcaggtccac
     2821 cctgacaccg gcatttcgtc caaggccatg ggcatcatga actcgtttgt gaacgacatt
     2881 ttcgagcgca tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc
     2941 acctccaggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac
     3001 gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagta atgaggcgcg
     3061 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
     3121 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3181 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3241 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3301 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     3361 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3421 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3481 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3541 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3601 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3661 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3721 tatcagcaca caattgccca ttatacgcgc gtataatgga ctattgtgtg ctgatatgat
     3781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     3841 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3901 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     3961 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4021 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4081 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4141 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4201 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     4261 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4321 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4381 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4441 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4501 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4561 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4621 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4681 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4741 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4801 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4861 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4921 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4981 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5041 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5101 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5161 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5221 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5281 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5341 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5401 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5461 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5521 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5581 attgatttaa aacttc
//
