LOCUS       sequence-470782-        5426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1313m 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 5426)
  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 5426)
  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: AiP14805 -
            pAAV-AiE1313m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    72..99
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    100..127
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      196..651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(283..302)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     493..514
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(668..687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     787..809
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(847..865)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        933..1037
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1038..1106,1107..1898)
                     /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(1256..1275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2069..2657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2558..2577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2714..2741
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2742..2769
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2776..2905
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3554..3563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3560..3562,3563..3565,3566..4276)
                     /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(3866..3885)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4213..4234
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4286..4342
                     /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             4355..4420
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4451..4828
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4915..4935)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5122..5233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   join(5351..5426,1..65)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5351..5426,1..54)
                     /label=AAV2 ITR
ORIGIN
        1 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
       61 gcaggggcgc ctatcagcac acaattgccc attatacgcg cgtataatgg actattgtgt
      121 gctgatatga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
      181 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      241 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
      301 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      361 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
      421 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
      481 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
      541 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
      601 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
      661 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
      721 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
      781 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
      841 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
      901 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      961 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1021 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1081 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1141 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1201 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1261 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1321 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1381 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1441 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1501 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1561 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1621 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1681 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1741 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1801 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1861 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1921 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1981 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2041 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2101 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2161 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2221 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2281 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2341 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2401 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2461 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2521 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2581 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2641 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2701 ggccttttgc tcatatcagc acacaattgc ccattatacg cgcgtataat ggactattgt
     2761 gtgctgataa catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     2821 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     2881 gccaactcca tcactagggg ttcctgcggc cgcacgcgtc accaagctgg catggtggct
     2941 gtcatttctg aaagtgatgt ggaaaattat aatcattatt ttcatcatcc actgagcctc
     3001 agaacaggct tcaagtctcc cccccccttt ttttaatctt ctcacgtttt tagtttattt
     3061 ttggaagtga gatcaatggg tcgtactttg ctttctttta ggaacctctg ggccacattt
     3121 ttaaaccaca cccagtggca gagcacgtga ctggaccttc atggaatatc tgatggtctt
     3181 gttgggcttg gtttcctgat cagctaatga tagccaggca gcacactctg ccagtcttgg
     3241 acattattat taagtccggt acatggtgga aataatgtgc taagtcataa taatatataa
     3301 caatgggaat gtgagctgta gcctctaata aggcatcagg tgcatgatag gtgaatattt
     3361 gtcaaaggac aaaacgtgct tacatggcta cctagagcaa tactacttgg ggaagtggtg
     3421 agtctatcat aggccctggg tgctttacaa agctttgttt gagctcgggc tgggcataaa
     3481 agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct
     3541 agcgctaccg gtcgccacca tggtctccaa gggcgaggag ctgttcaccg gggtggtgcc
     3601 catcctggtc gagctggacg gcgatgtcaa cggccacaag ttcagcgtgt ccggcgaggg
     3661 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     3721 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg
     3781 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3841 ccaggagcgc accatcttct tcaaagacga cggcaactac aagacccgcg ccgaggtgaa
     3901 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3961 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     4021 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     4081 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4141 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     4201 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4261 ggacgagctg tacaagggct ctggtgctac caacttctca ctgttgaaac aggcagggga
     4321 tgtagaggag aatccagggc ctggtgctag tggagactac aaagaccatg acggagatta
     4381 taaagatcat gacatcgatt acaaggatga cgatgacaag tccggactca gatctggagg
     4441 ctccggaggc ccagagccag cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc
     4501 ggtgactaag gcgcagaaga aaggcggcaa gaagcgcaag cgcagccgca aggagagcta
     4561 ttccatctat gtgtataagg ttctgaagca ggtccaccct gacaccggca tttcgtccaa
     4621 ggccatgggc atcatgaact cgtttgtgaa cgacattttc gagcgcatcg caggtgaggc
     4681 ttcccgcctg gcgcattaca acaagcgctc gaccatcacc tccagggaga tccagacggc
     4741 cgtgcgcctg ctgctgcctg gggagttggc caagcacgcc gtgtccgagg gtactaaggc
     4801 catcaccaag tacaccagcg ctaagtaatg aggcgcgccg cggccgcgaa ttcgatatca
     4861 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4921 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4981 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     5041 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5101 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5161 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5221 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5281 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     5341 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     5401 ctcactgagg ccgggcgacc aaaggt
//
