LOCUS       sequence-470753-        5200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1389h 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 5200)
  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 5200)
  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: AiP14979 -
            pAAV-AiE1389h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      83..538
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(170..189)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     380..401
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(555..574)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     674..696
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(734..752)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        820..924
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(925..993,994..1785)
                     /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(1143..1162)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1956..2544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2445..2464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2601..2628
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2629..2656
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2663..2792
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3215..3224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3221..3223,3224..3226,3227..3937)
                     /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(3527..3546)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3874..3895
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3947..4003
                     /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             4016..4081
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4112..4489
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4576..4596)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4783..4894
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5012..5152
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5012..5141
                     /label=AAV2 ITR
     misc_feature    5159..5186
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    join(5187..5200,1..14)
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 attgtgtgct gatatgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
       61 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      121 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
      181 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      241 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
      301 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      361 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
      421 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
      481 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
      541 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
      601 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
      661 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
      721 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
      781 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      841 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      901 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      961 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1021 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1081 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1141 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1201 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1261 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1321 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1381 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1441 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1501 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1561 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1621 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1681 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1741 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1801 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1861 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1921 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1981 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2041 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2101 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2161 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2221 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2281 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2341 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2401 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2461 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2521 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2581 ttttgctggc cttttgctca tatcagcaca caattgccca ttatacgcgc gtataatgga
     2641 ctattgtgtg ctgataacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     2701 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     2761 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtcatc ctctcctctt
     2821 gccttgccta ggccatggca gtagttttct ccatcatctc actcccacag cctacccttc
     2881 tgcagttctc ttcacactgt tgctactcaa ctcttctaaa agcagaatat gatcatctta
     2941 atgctttgct tgaaacacaa tgagctgttc ttagctacag agtaaaatcc aaactctagc
     3001 atctggctcc agttcactat tgcatttcca ttttagggaa ctgcgaagct tctcaagcat
     3061 caaatttcac atgaatgaag gggagtagga tcttgttaaa atgcaaatcc tggatctggc
     3121 tgagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     3181 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtctcca agggcgagga
     3241 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgatgtca acggccacaa
     3301 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     3361 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     3421 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3481 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaagacg acggcaacta
     3541 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3601 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3661 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     3721 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     3781 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     3841 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3901 cgccgggatc actctcggca tggacgagct gtacaagggc tctggtgcta ccaacttctc
     3961 actgttgaaa caggcagggg atgtagagga gaatccaggg cctggtgcta gtggagacta
     4021 caaagaccat gacggagatt ataaagatca tgacatcgat tacaaggatg acgatgacaa
     4081 gtccggactc agatctggag gctccggagg cccagagcca gcgaagtctg ctcccgcccc
     4141 gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca agaagcgcaa
     4201 gcgcagccgc aaggagagct attccatcta tgtgtataag gttctgaagc aggtccaccc
     4261 tgacaccggc atttcgtcca aggccatggg catcatgaac tcgtttgtga acgacatttt
     4321 cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac aacaagcgct cgaccatcac
     4381 ctccagggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc
     4441 cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaagtaat gaggcgcgcc
     4501 gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac
     4561 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4621 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4681 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4741 gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc
     4801 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4861 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4921 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4981 atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     5041 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     5101 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgccta
     5161 tcagcacaca attgcccatt atacgcgcgt ataatggact
//
