LOCUS       sequence-449641-        5541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1367m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical L2/3_IT neurons (note HPF dentate 
            gyrus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5541)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5541)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP14937 -
            pAAV-AiE1367m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5541
                     /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      894..903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(900..902,903..905,906..1616)
                     /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(1206..1225)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1553..1574
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1626..1682
                     /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             1695..1760
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1791..2168
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2255..2275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2462..2573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2691..2831
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2691..2820
                     /label=AAV2 ITR
     misc_feature    2838..2865
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2866..2893
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2962..3417
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3049..3068)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3259..3280
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3434..3453)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3553..3575
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3613..3631)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3699..3803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3804..3872,3873..4664)
                     /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(4022..4041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4835..5423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5324..5343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5480..5507
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5508..5535
                     /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 gcgtatttca ttaaaatgaa cccagactgt aaacttgacc
      181 tcaccaagtc atttttcttc aataataaag ttttccagct atatttactt atagtttgct
      241 cccacatatc tgaaagctgc catctgtgga taataagaac aaagctttat cacagattgt
      301 tttaatttcc tgcctgctat cagcaccaag aaggtaaccc tggaaacggg gttcatagca
      361 tcctccttcc tgctgcctgt tagtcacaac cacaaaggaa accagaacct cctgaagttg
      421 gtctgtccac accaccactg cccttgtcag ggttaagcaa ggaagcaaca cggttgagca
      481 gccatcccgt aagcctcttg cccagtagga ctcctcctga aaatatagta cagcacacat
      541 acgatgctct ctaagcctct tctgaagcaa ggatgtcaga cccttccgtg gggcccagcc
      601 acagtctctg tgggtctggg tgggcttgag aatctgattt ctgaaaacgc ctagacactg
      661 tgatgctggt ccaccaccac tgagagcctg ctgtggagca tctcagttgc tacccagaac
      721 cttcacatcg cctactgttc ctgcccacat atgtatgtgc acatttagta caaacagacc
      781 tgttttgcca gggagaaggg gagctcgggc tgggcataaa agtcagggca gagccatcta
      841 ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca
      901 tggtctccaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      961 gcgatgtcaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1021 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1081 tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
     1141 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1201 tcaaagacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1261 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1321 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     1381 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     1441 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1501 acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1561 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggct
     1621 ctggtgctac caacttctca ctgttgaaac aggcagggga tgtagaggag aatccagggc
     1681 ctggtgctag tggagactac aaagaccatg acggagatta taaagatcat gacatcgatt
     1741 acaaggatga cgatgacaag tccggactca gatctggagg ctccggaggc ccagagccag
     1801 cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga
     1861 aaggcggcaa gaagcgcaag cgcagccgca aggagagcta ttccatctat gtgtataagg
     1921 ttctgaagca ggtccaccct gacaccggca tttcgtccaa ggccatgggc atcatgaact
     1981 cgtttgtgaa cgacattttc gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca
     2041 acaagcgctc gaccatcacc tccagggaga tccagacggc cgtgcgcctg ctgctgcctg
     2101 gggagttggc caagcacgcc gtgtccgagg gtactaaggc catcaccaag tacaccagcg
     2161 ctaagtaatg aggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca
     2221 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2281 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2341 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     2401 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg
     2461 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2521 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2581 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2641 tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct
     2701 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2761 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2821 ctgcctgcag gggcgcctat cagcacacaa ttgcccatta tacgcgcgta taatggacta
     2881 ttgtgtgctg atatgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2941 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3001 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     3061 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3121 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3181 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3241 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     3301 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     3361 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     3421 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3481 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3541 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3601 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3661 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3721 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3781 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3841 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3901 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3961 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4021 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4081 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4141 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4201 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4261 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4321 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4381 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4441 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4501 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4561 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4621 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4681 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4741 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4801 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4861 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4921 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4981 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5041 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5101 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5161 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5221 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5281 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5341 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5401 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5461 tttgctggcc ttttgctcat atcagcacac aattgcccat tatacgcgcg tataatggac
     5521 tattgtgtgc tgataacatg t
//
