LOCUS       sequence-461891-        5359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1388h 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 5359)
  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 5359)
  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: AiP14978 -
            pAAV-AiE1388h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      712..721
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(718..720,721..723,724..1434)
                     /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(1024..1043)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1371..1392
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1444..1500
                     /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             1513..1578
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1609..1986
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2073..2093)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2280..2391
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2509..2649
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2509..2638
                     /label=AAV2 ITR
     misc_feature    2656..2683
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2684..2711
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2780..3235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2867..2886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3077..3098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3252..3271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3371..3393
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3431..3449)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3517..3621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3622..3690,3691..4482)
                     /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(3840..3859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4653..5241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5142..5161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5298..5325
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5326..5353
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagtcac atttgcaggt atatgtgtgt ggtttctgtc
      181 ttccacccct ctcctccttt gtttggttta accagctcgc tgtgatgtca agaacctcac
      241 actaattcct tagggttctc cgatatgaag tcaatgtgct tgccttccgg gggatatgca
      301 ttattttagg actgttctat tctgacactt tatcatcact gtaataatca tcagtgtaca
      361 gacatgccag ctagcaaggt aggcacttgg gaagaattgt gaaactgcaa ttaccaccag
      421 gtgcctccta caagtatttc aggaactgtt agaactctct ctgcttgaaa ctgggaacac
      481 ttggcagtcc tgcttagctt tacaaatctg ccaagtttgg taaatttagt ttcgacattt
      541 aaacagcatg gtagatttac atcaatttaa aaaaaaaact ggctgaattt ctgtatgacc
      601 ggccactgtt ttcaatagga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      661 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
      721 gtctccaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      781 gatgtcaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      841 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      901 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      961 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1021 aaagacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1081 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1141 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1201 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1261 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1321 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1381 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggctct
     1441 ggtgctacca acttctcact gttgaaacag gcaggggatg tagaggagaa tccagggcct
     1501 ggtgctagtg gagactacaa agaccatgac ggagattata aagatcatga catcgattac
     1561 aaggatgacg atgacaagtc cggactcaga tctggaggct ccggaggccc agagccagcg
     1621 aagtctgctc ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa
     1681 ggcggcaaga agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtataaggtt
     1741 ctgaagcagg tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaactcg
     1801 tttgtgaacg acattttcga gcgcatcgca ggtgaggctt cccgcctggc gcattacaac
     1861 aagcgctcga ccatcacctc cagggagatc cagacggccg tgcgcctgct gctgcctggg
     1921 gagttggcca agcacgccgt gtccgagggt actaaggcca tcaccaagta caccagcgct
     1981 aagtaatgag gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     2041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2161 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     2221 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     2281 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2341 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2401 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2461 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2521 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2581 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2641 gcctgcaggg gcgcctatca gcacacaatt gcccattata cgcgcgtata atggactatt
     2701 gtgtgctgat atgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2761 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2821 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     2881 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2941 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     3001 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3061 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     3121 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     3181 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     3241 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     3301 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3361 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3421 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3481 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3541 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3601 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3661 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3721 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3781 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3841 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3901 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3961 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4021 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4081 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4141 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4201 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4261 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4321 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4381 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4441 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4501 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4561 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4621 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4681 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4741 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4801 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4861 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4921 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4981 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5041 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5101 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5161 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5221 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5281 tgctggcctt ttgctcatat cagcacacaa ttgcccatta tacgcgcgta taatggacta
     5341 ttgtgtgctg ataacatgt
//
