LOCUS       sequence-470612-        5305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0944m 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 5305)
  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 5305)
  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: AiP13297 -
            pAAV-AiE0944m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   458..587
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(649..1237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(729..748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1408..2199,2200..2268))
                     /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     2031..2050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2269..2373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2441..2459
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2497..2519)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2619..2638
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2655..3110)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2792..2813)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3004..3023
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3185..3325
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3196..3325
                     /label=AAV2 ITR
     polyA_signal    3443..3554
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3741..3761
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3848..4225)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(4256..4321)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4334..4390)
                     /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             complement(join(4400..5110,5111..5113,5114..5116))
                     /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(4442..4463)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4791..4810
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      5113..5122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caattgtccc gagtggcagt aggccttcca aaaggcagga ggagctgtgg gctgaggttt
       61 ggagtgcaga tcttcgactg cacgtagacg tgtggactgg aagatggagc tggttgtcct
      121 gggtgggagc aggcttccca ggaaacaggc agaactgtgg gctgatgagt ggggtgcaga
      181 tccaagattg caagtagagg tgtggaccag aaggaagttg gagctctgga aggcgggggc
      241 agacctcaca gctgctaggc ttgctggggt cccagcaggc aggatattgg ggcatggatt
      301 tctcacctgg tgttccaggt gggagcaggc cttccaagag gtggacagag ttgtgggctg
      361 gggagtaggg tgccaacctg cagttggttc ctctttttcc ttagtattaa ttatcattga
      421 gtctgttaag tctctgggga ggtacgcgtg cggccgcagg aacccctagt gatggagttg
      481 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
      541 cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa
      601 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      661 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      721 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      781 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      841 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      901 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      961 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1021 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1081 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1141 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1201 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1261 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1321 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1381 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1441 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1501 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1561 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1621 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1681 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     1741 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1801 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1861 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1921 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1981 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2041 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2101 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2161 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2221 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2281 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2341 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     2401 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     2461 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2521 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2581 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     2641 gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     2701 ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa
     2761 agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2821 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2881 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2941 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3001 ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct
     3061 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg
     3121 ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     3181 cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
     3241 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     3301 gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct
     3361 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
     3421 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
     3481 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     3541 actagaaggc acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag
     3601 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc
     3661 aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag
     3721 gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca
     3781 atctttcaca aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg
     3841 cgcctcatta cttagcgctg gtgtacttgg tgatggcctt agtaccctcg gacacggcgt
     3901 gcttggccaa ctccccaggc agcagcaggc gcacggccgt ctggatctcc cgggaggtga
     3961 tggtcgagcg cttgttgtaa tgcgccaggc gggaagcctc tcctgcgatg cgctcgaaaa
     4021 tgtcgttcac aaacgaattc atgatgccca tggccttgga cgaaatgccg gtgtcagggt
     4081 ggacctgctt cagaaccttg tacacataga tggaatagct ctccttgcgg ctgcgcttgc
     4141 gcttcttgcc gcctttcttc tgcgccttag tcaccgcctt cttggagccc tttttcgggg
     4201 cgggagcaga cttcgctggc tctgggcctc cggagcctcc agatctgagt ccggacttgt
     4261 catcgtcatc cttgtaatcg atgtcatgat ctttataatc tccgtcatgg tctttgtagt
     4321 ctccactagc accaggccct ggattctcct ctacatcccc tgcctgtttc aacagtgaga
     4381 agttggtagc accagagccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     4441 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg
     4501 actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     4561 tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga
     4621 agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt
     4681 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     4741 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     4801 tgccgtcgtc tttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     4861 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc
     4921 ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct
     4981 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     5041 ggccgttgac atcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     5101 cgcccttgga gaccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga
     5161 agcaaatgta agcaatagat ggctctgccc tgacttttat gcccagcccg agctctgagt
     5221 gtgttcttgc acttgccttt ggccatctgg ttgtctctga tgctggttgg cctggtagtt
     5281 cccgggagta gcaagcctct gggac
//
