LOCUS       sequence-460663-        5053 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV vector for pan-neuronal expression of nuclear mTFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5053)
  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 5053)
  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: AiP14019 - pAAV-hSyn1-mTFP1-10aa-H2B-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..5053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        183..630
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      666..675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             672..1379
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             1410..1787
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1880..1900)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2087..2198
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2316..2456
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2316..2445
                     /label=AAV2 ITR
     rep_origin      2531..2986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2618..2637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2828..2849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3003..3022)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3122..3144
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3182..3200)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3268..3372
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3373..3441,3442..4233)
                     /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(3591..3610)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4404..4992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4893..4912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aagtgtctag actgcagagg gccctgcgta
      181 tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc
      241 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca
      301 gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg
      361 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc
      421 gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc
      481 cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg
      541 cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc
      601 ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggctagagga tccttcgaag
      661 ctagcgccac catggtgagc aagggcgagg agaccacaat gggcgtaatc aagcccgaca
      721 tgaagatcaa gctgaagatg gagggcaacg tgaatggcca cgccttcgtg atcgagggcg
      781 agggcgaggg caagccctac gacggcacca acaccatcaa cctggaggtg aaggagggag
      841 cccccctgcc cttctcctac gacattctga ccaccgcgtt cgcctacggc aacagggcct
      901 tcaccaagta ccccgacgac atccccaact acttcaagca gtccttcccc gagggctact
      961 cttgggagcg caccatgacc ttcgaggaca agggcatcgt gaaggtgaag tccgacatct
     1021 ccatggagga ggactccttc atctacgaga tacacctcaa gggcgagaac ttccccccca
     1081 acggccccgt gatgcagaag aagaccaccg gctgggacgc ctccaccgag aggatgtacg
     1141 tgcgcgacgg cgtgctgaag ggcgacgtca agcacaagct gctgctggag ggcggcggcc
     1201 accaccgcgt tgacttcaag accatctaca gggccaagaa ggcggtgaag ctgcccgact
     1261 atcactttgt ggaccaccgc atcgagatcc tgaaccacga caaggactac aacaaggtga
     1321 ccgtttacga gagcgccgtg gcccgcaact ccaccgacgg catggacgag ctgtacaagt
     1381 ccggactcag atctggaggc tccggaggcc cagagccagc gaagtctgct cccgccccga
     1441 aaaagggctc caagaaggcg gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc
     1501 gcagccgcaa ggagagctat tccatctatg tgtataaggt tctgaagcag gtccaccctg
     1561 acaccggcat ttcgtccaag gccatgggca tcatgaactc gtttgtgaac gacattttcg
     1621 agcgcatcgc aggtgaggct tcccgcctgg cgcattacaa caagcgctcg accatcacct
     1681 ccagggagat ccagacggcc gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg
     1741 tgtccgaggg tactaaggcc atcaccaagt acaccagcgc taagtaatga gtcgacggcg
     1801 cgcccctgca gggaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1861 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1921 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1981 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2041 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     2101 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2161 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2221 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2281 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     2341 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2401 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2461 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2521 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2581 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     2641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     2881 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     2941 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     3001 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3061 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3121 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3181 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3241 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3301 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3361 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3421 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3481 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3541 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3601 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3661 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3721 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3781 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3841 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3901 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3961 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4021 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4081 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4141 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4201 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4261 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4321 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4381 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4441 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4501 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4561 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4621 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4681 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4741 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4801 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4861 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4921 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4981 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5041 tttgctcaca tgt
//
