LOCUS       sequence-470599-        5521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0344m 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 5521)
  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 5521)
  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: AiP14537 -
            pAAV-AiE0344m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    120..147
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    148..175
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      244..699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(331..350)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     541..562
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(716..735)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     835..857
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(895..913)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        981..1085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1086..1154,1155..1946)
                     /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(1304..1323)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2117..2705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2606..2625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2762..2789
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2790..2817
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2824..2953
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3697..3706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3703..3705,3706..3708,3709..4419)
                     /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(4009..4028)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4356..4377
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4429..4485
                     /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             4498..4563
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4594..4971
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(5058..5078)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5265..5376
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   join(5494..5521,1..113)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5494..5521,1..102)
                     /label=AAV2 ITR
ORIGIN
        1 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
       61 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
      121 atcagcacac aattgcccat tatacgcgcg tataatggac tattgtgtgc tgatatgatg
      181 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
      241 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      301 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
      361 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      421 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
      481 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      541 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
      601 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
      661 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
      721 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
      781 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
      841 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
      901 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
      961 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1021 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1081 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1141 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1201 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1261 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1321 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1381 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1441 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1501 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1561 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1621 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1681 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1741 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1801 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1861 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1921 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1981 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2041 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2101 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2161 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2221 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2281 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2341 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2401 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2461 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2521 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2581 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2641 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2701 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2761 atatcagcac acaattgccc attatacgcg cgtataatgg actattgtgt gctgataaca
     2821 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     2881 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2941 actaggggtt cctgcggccg cacgcgtagt caggggtcag tcctacttat ggcctgacag
     3001 cttgtcacag cccccacaac ccgaatgtct ccttaaacct ggcctccaca gctcagcatc
     3061 gccaacactg acagtgtcaa agttgctggt ttccggagag gaaggcagag aggagggatg
     3121 agcaaaagaa caaagacccc gccctgtgcc tgcacctgca tcacccagaa ctctctgaaa
     3181 tcatcagctc tctgaaatcc ccgtgtctct gagtcccctt gagcagggag tagaccgcat
     3241 gcagctgagt ccacggagac tctgaagagc acaggtggca cgagacagga atgaaaggac
     3301 aggagccctt agccacggaa tttcaccttt tctctctggc tcacattgcc atgaaggagg
     3361 tttatgctca atgattgtag ctttagaatt cacagctcca ctaccggtgg aaatagttgc
     3421 atttctttct gtagcccagg ctggcttcaa actcattatg aacttgaact cctgacctcc
     3481 cttcctccac ccaccaagtt ctggaattac agccatggaa aaccaccatg cccagttgag
     3541 aaatacctgt gtaactattg tttttgatac aagtattata attggggata ggaagataga
     3601 ggagagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
     3661 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtctc caagggcgag
     3721 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgatgt caacggccac
     3781 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3841 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     3901 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     3961 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaaaga cgacggcaac
     4021 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4081 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4141 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     4201 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     4261 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     4321 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4381 gccgccggga tcactctcgg catggacgag ctgtacaagg gctctggtgc taccaacttc
     4441 tcactgttga aacaggcagg ggatgtagag gagaatccag ggcctggtgc tagtggagac
     4501 tacaaagacc atgacggaga ttataaagat catgacatcg attacaagga tgacgatgac
     4561 aagtccggac tcagatctgg aggctccgga ggcccagagc cagcgaagtc tgctcccgcc
     4621 ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc
     4681 aagcgcagcc gcaaggagag ctattccatc tatgtgtata aggttctgaa gcaggtccac
     4741 cctgacaccg gcatttcgtc caaggccatg ggcatcatga actcgtttgt gaacgacatt
     4801 ttcgagcgca tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc
     4861 acctccaggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac
     4921 gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagta atgaggcgcg
     4981 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
     5041 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5101 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5161 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5221 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     5281 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     5341 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     5401 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     5461 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     5521 c
//
