LOCUS       sequence-470769-        5435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0893m 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 5435)
  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 5435)
  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: AiP14414 -
            pAAV-AiE0893m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    77..104
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    105..132
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      201..656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(288..307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     498..519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(673..692)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     792..814
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(852..870)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        938..1042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1043..1111,1112..1903)
                     /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(1261..1280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2074..2662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2563..2582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2719..2746
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2747..2774
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2781..2910
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3568..3577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3574..3576,3577..3579,3580..4290)
                     /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(3880..3899)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4227..4248
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4300..4356
                     /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             4369..4434
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4465..4842
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4929..4949)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5136..5247
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   join(5365..5435,1..70)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5365..5435,1..59)
                     /label=AAV2 ITR
ORIGIN
        1 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
       61 tgcctgcagg ggcgcctatc agcacacaat tgcccattat acgcgcgtat aatggactat
      121 tgtgtgctga tatgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      181 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
      241 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
      301 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
      361 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
      421 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
      481 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
      541 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
      601 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
      661 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
      721 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
      781 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
      841 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
      901 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      961 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1021 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1081 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1141 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1201 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1261 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1321 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1381 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1441 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1501 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1561 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1621 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1681 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1741 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1801 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1861 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1921 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1981 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2041 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2101 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2161 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2221 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2281 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2341 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2401 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2461 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2521 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2581 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2641 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2701 ttgctggcct tttgctcata tcagcacaca attgcccatt atacgcgcgt ataatggact
     2761 attgtgtgct gataacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     2821 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2881 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtgaaaac aatttctgtg
     2941 ctgacaacag agagggcatg agattctaat tcttctggtc aaagttttta tgttcatttt
     3001 taagttaaat tgttgaatgc cttttaaaaa tgtagctgtc agagatattt tctcattagt
     3061 gtctctagag aagtttagga aacactactc atgagaagta ggtcaatggg cagctgccta
     3121 tcacagcctc atgagaagct agcaagtgtt ttaggacaag gcccagaata gacagatgct
     3181 cccatgcctt cagtcaattg cagggctatc ccaatttgcc tgctagggct aaggaggaca
     3241 ggcatgcatt ttctgggaga atgtagagga accagaccca aagaacaagc tctgctgctg
     3301 cgcccaataa tcattagaac cagcaggttg aaaactacag gccttggagt aagaacaaat
     3361 agcttgtgac tctccctgta tggaatggta tatttttaat ttttttaaaa gatttattta
     3421 tttcatttat atgagtacac tgttgctgtc ttaagacaca ccagaagagg acatgagctc
     3481 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
     3541 gatctttcga agctagcgct accggtcgcc accatggtct ccaagggcga ggagctgttc
     3601 accggggtgg tgcccatcct ggtcgagctg gacggcgatg tcaacggcca caagttcagc
     3661 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     3721 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
     3781 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3841 cccgaaggct acgtccagga gcgcaccatc ttcttcaaag acgacggcaa ctacaagacc
     3901 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3961 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     4021 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     4081 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     4141 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     4201 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     4261 atcactctcg gcatggacga gctgtacaag ggctctggtg ctaccaactt ctcactgttg
     4321 aaacaggcag gggatgtaga ggagaatcca gggcctggtg ctagtggaga ctacaaagac
     4381 catgacggag attataaaga tcatgacatc gattacaagg atgacgatga caagtccgga
     4441 ctcagatctg gaggctccgg aggcccagag ccagcgaagt ctgctcccgc cccgaaaaag
     4501 ggctccaaga aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc
     4561 cgcaaggaga gctattccat ctatgtgtat aaggttctga agcaggtcca ccctgacacc
     4621 ggcatttcgt ccaaggccat gggcatcatg aactcgtttg tgaacgacat tttcgagcgc
     4681 atcgcaggtg aggcttcccg cctggcgcat tacaacaagc gctcgaccat cacctccagg
     4741 gagatccaga cggccgtgcg cctgctgctg cctggggagt tggccaagca cgccgtgtcc
     4801 gagggtacta aggccatcac caagtacacc agcgctaagt aatgaggcgc gccgcggccg
     4861 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4921 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4981 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     5041 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5101 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     5161 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     5221 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     5281 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     5341 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     5401 tgcgcgctcg ctcgctcact gaggccgggc gacca
//
