LOCUS       sequence-466257-        4994 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2172m 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 4994)
  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 4994)
  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: AiP1401 - pAAV-AiE2172m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     412..431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   573..702
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1578..1587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1584..1586,1587..1589,1590..2300)
                     /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(1629..1650)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1890..1909)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2237..2258
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2393..2413)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2600..2711
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2829..2969
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2829..2958
                     /label=AAV2 ITR
     rep_origin      3044..3499
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3131..3150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3341..3362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3516..3535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3635..3657
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3695..3713)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3781..3885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3886..3954,3955..4746)
                     /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(4104..4123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4917..4994,1..511)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
       61 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      541 cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct
      601 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
      661 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgttgta
      721 agagtaataa tcgccggcgc ttagatatgc aagttcctag ccctctgttt aaatttcaat
      781 ttcactttag ggttctgaga tggacttgaa gtggtcactg gatctgcctc ttcccaatgt
      841 gaccacaatg agcaaatctc ctttctggtt ctctctattg gtctggttgg gtagatctat
      901 ttatctattg tcttcttggg gtagagactg tgacaggact gtgacccccc cccatctttt
      961 ggtagcagtc cctgctcttt gagagtccca ggagatgagc tcagctcggc cccggaggtg
     1021 atgtcatctg agtcttctcc ggagggactt ccaagggagc cctgtgtgag ttcactctgc
     1081 acttctgttt tcaaacagct gctctctgct ccggtctctg ggtgccactc tcagttctct
     1141 ctttgattcc cacccaaacc aacagagaag ccaagtgaaa gagtaaaaac tcccttatgt
     1201 atgtaatcaa tccactgaaa acgaaggtcc aaggagaagc cagcgtgcct gtctcgggga
     1261 cctacactcc agcatacaca aggacattgc tacctttaaa gtcatgttga ccaccttgcc
     1321 ccaagggtct catggtagcg catcaacccc aaagggactc actgttcact tcttccccct
     1381 ccgtcctcac tgttctcatt ggtttgtttt tctaagtgta tgagtgcctg gcctgcatgt
     1441 atgtctagtg ggcgtaatgc ccacagaggc cacaagcgcc ggcggagctc gggctgggca
     1501 taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga
     1561 agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1621 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1681 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
     1741 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg
     1801 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1861 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1921 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1981 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2041 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     2101 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2161 ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca
     2221 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2281 gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata
     2341 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2401 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2461 tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca
     2521 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2581 tggctcgaga gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2641 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2701 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2761 gacagcaagg gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga
     2821 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     2881 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     2941 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3001 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3061 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3121 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3181 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3241 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3301 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3361 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     3421 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3481 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3541 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3601 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3661 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     3721 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3781 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3841 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3901 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3961 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4021 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4081 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4141 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4201 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4261 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4321 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4381 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4441 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4501 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4561 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4621 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4681 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4741 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4801 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4861 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4921 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4981 gtggtttgtt tgcc
//
