LOCUS       sequence-466201-        5877 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2036m 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 5877)
  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.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: AiP1173 - pAAV-AiE2036m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      129..717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     618..637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   779..908
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1453..1462
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1459..1464,1465..1485,1486..2757)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    2782..3370
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2835..2855)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3253..3264)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3402..3878
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3601..3620)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3918..4058
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3918..4047
                     /label=AAV2 ITR
     rep_origin      4133..4588
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4220..4239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4430..4451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4605..4624)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4724..4746
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4784..4802)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4870..4974
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4975..5043,5044..5835)
                     /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(5193..5212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
       61 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      121 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      181 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      241 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      301 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      361 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      421 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      481 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      541 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      601 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      661 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      721 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc
      781 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg
      841 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
      901 gggttcctgc ggccgcacgc gtgagtaata ggcagatctc tataccagcc taacttctcc
      961 ctaccacccc cctccatctt ccttcttctt gcagtgcaaa cgccaagtca tatggacttg
     1021 gattccgact ctcttttctt tgccaagact catattattg agcaagtgag gagagttgag
     1081 ccaaactgaa ctaagtgacg ccacagcaca ctgtgtagtg cagagctagc actttggata
     1141 ttgtgtaatt aaggctttaa tgaacatcac atagagaatt agactgcaga ttgtgacaca
     1201 aataaatcta cttggctttt agaaacattc cttaaaaaga aatattttgg taccagaatg
     1261 tgttcaggag ggaaagagcc tgtgcccttg acacaggaag tagataacac ctgtcagaag
     1321 aatcgaaacc acctgcttgg gtgcatctgg taataagttt gcagctggtg ctacactcta
     1381 cccttaagga gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     1441 gcttctggcg tggccaccat ggctcctaag aagaagagga aggtgatgag ccagttcgac
     1501 atcctgtgca agaccccccc caaggtgctg gtgcggcagt tcgtggagag attcgagagg
     1561 cccagcggcg agaagatcgc cagctgtgcc gccgagctga cctacctgtg ctggatgatc
     1621 acccacaacg gcaccgccat caagagggcc accttcatga gctacaacac catcatcagc
     1681 aacagcctga gcttcgacat cgtgaacaag agcctgcagt tcaagtacaa gacccagaag
     1741 gccaccatcc tggaggccag cctgaagaag ctgatccccg cctgggagtt caccatcatc
     1801 ccttacaacg gccagaagca ccagagcgac atcaccgaca tcgtgtccag cctgcagctg
     1861 cagttcgaga gcagcgagga ggccgacaag ggcaacagcc acagcaagaa gatgctgaag
     1921 gccctgctgt ccgagggcga gagcatctgg gagatcaccg agaagatcct gaacagcttc
     1981 gagtacacca gcaggttcac caagaccaag accctgtacc agttcctgtt cctggccaca
     2041 ttcatcaact gcggcaggtt cagcgacatc aagaacgtgg accccaagag cttcaagctg
     2101 gtgcagaaca agtacctggg cgtgatcatt cagtgcctgg tgaccgagac caagacaagc
     2161 gtgtccaggc acatctactt tttcagcgcc agaggcagga tcgaccccct ggtgtacctg
     2221 gacgagttcc tgaggaacag cgagcccgtg ctgaagagag tgaacaggac cggcaacagc
     2281 agcagcaaca agcaggagta ccagctgctg aaggacaacc tggtgcgcag ctacaacaag
     2341 gccctgaaga agaacgcccc ctaccccatc ttcgctatca agaacggccc taagagccac
     2401 atcggcaggc acctgatgac cagctttctg agcatgaagg gcctgaccga gctgacaaac
     2461 gtggtgggca actggagcga caagagggcc tccgccgtgg ccaggaccac ctacacccac
     2521 cagatcaccg ccatccccga ccactacttc gccctggtgt ccaggtacta cgcctacgac
     2581 cccatcagca aggagatgat cgccctgaag gacgagacca accccatcga ggagtggcag
     2641 cacatcgagc agctgaaggg cagcgccgag ggcagcatca gataccccgc ctggaacggc
     2701 atcatcagcc aggaggtgct ggactacctg agcagctaca tcaacaggcg gatctgagaa
     2761 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2821 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2881 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2941 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3001 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3061 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3121 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3181 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3241 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3301 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3361 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     3421 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3481 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3541 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     3601 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3661 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3721 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3781 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3841 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     3901 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3961 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4021 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4081 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4141 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4201 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4261 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4321 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4381 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4441 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4501 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4561 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4621 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4681 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4741 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4801 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4861 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4921 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4981 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5041 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5101 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5161 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5221 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5281 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5341 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5401 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5461 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5521 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5581 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5641 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5701 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5761 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5821 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgattta
//
