LOCUS       sequence-446472-        5333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 AAV virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5333)
  AUTHORS   Kussick E, Johansen N, Taskin N, Chowdhury A, Quinlan MA, Fraser A, 
            Clark AG, Wynalda B, Martinez R, Groce EL, Reding M, Liang E, Shulga
            L, Huang C, Casper T, Clark M, Ho W, Gao Y, van Velthoven CTJ, 
            Sobieski C, Ferrer R, Berg MR, Curtis BC, English C, Day JC, Fortuna
            MG, Donadio N, Newman D, Yao S, Chakka AB, Goldy J, Torkelson A, 
            Guzman JB, Chakrabarty R, Nguy B, Guilford N, Pham TH, Wright V, 
            Ronellenfitch K, Naidoo R, Kenney J, Williford A, Ramakrishnan C, 
            Drinnenberg A, Gudsnuk K, Thyagarajan B, Smith KA, Dee N, Deisseroth
            K, Zeng H, Yao Z, Tasic B, Levi BP, Hodge R, Bakken TE, Lein ES, 
            Ting JT, Daigle TL
  TITLE     Enhancer AAVs for targeting spinal motor neurons and descending 
            motor pathways in rodents and macaque.
  JOURNAL   Cell Rep. 2025 Jun 24;44(6):115730. doi: 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21.
  PUBMED    40403722
REFERENCE   2  (bases 1 to 5333)
  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 Rep.";
            date: "2025-06-24"; volume: "44(6):115730. doi"; pages: " 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21"
COMMENT     Sequence Label: HCT6_rAAV-eHGT_1141m-minBglobin-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5333
                     /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      1345..1354
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1351..1353,1354..1356,1357..2067)
                     /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(1396..1417)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1657..1676)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2004..2025
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2160..2180)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2367..2478
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2596..2736
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2596..2725
                     /label=AAV2 ITR
     rep_origin      2811..3266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2898..2917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3108..3129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3283..3302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3402..3424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3462..3480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3653..3721,3722..4513)
                     /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(3871..3890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4684..5272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5173..5192
                     /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 gcgttgtaag agtaataatc gccggcggag ttattggtta
      181 tgaagccaaa cccataaaac tggtggtttg gagaaggacg aaaaggctca gaaaaccaac
      241 agttcctatt ctgaactcgg gccaggaatg gctgaacact catgtaggct cttcaaaggc
      301 aagaaaacaa acaaatgttg aaaaaccact actgcttcct ctgaggattc ccttgcgggg
      361 gctgcgtaca cctcccaagg ccagactgac gatctgaaag agagagattt ggctattagt
      421 ctccacaaac tggtctgttc ctagatggcg ctgatgttaa agtgcaaaga tcctttaagt
      481 gtagattctt ttgagaaggg gatggtatta aggacatttc attggtctgt ggtccttggt
      541 cactgtgtcc agtcagcaaa atggtcccag ttgctgatct ctggctggca ggccaaattc
      601 tggccaaaat gctcaccgca ctgtggacag agctggcatg cttacaactt ggatgtccct
      661 tccgggacta aattcccaat attggtcaga tgtgaaaagt actgggactt ctgcccagaa
      721 aatacttcag gagtgacaag taattagagg caggcttggg acaattattt ataacttggg
      781 acacttccac attaaagact ccaccatctt ctcaagagag actggccaga gcactcggct
      841 ctgctttatt tgccagcaag attcaccagg ctatgtcctc tgccattata gtcttttgaa
      901 aagaggggat aatatatttg tctttaaaac gtcattagtt atttcttctt tgctttgact
      961 tgtccaacta ctcctcttgc ttgtacaggc cagttctgtg tgtacagttt tctatactta
     1021 acaacttgtt tggtctgttc ttttggaaaa atagttacac agtgtccagt gcagatatta
     1081 gcattggcct tagctagtta catagtagcg catggccggg agaggtttgt ttgagcttgt
     1141 ttgagccgtc cccattgcat gaaaggctgt ttagtcttcc ctgagctcag gaaggaccat
     1201 agttaggaag tgagtcaaca tcccgcagca tcccacatgg acccgccggc ggagctcggg
     1261 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
     1321 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     1381 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1441 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1501 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1561 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1621 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1681 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1741 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1801 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1861 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1921 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1981 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2041 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     2101 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2161 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2221 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     2281 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2341 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     2401 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2461 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2521 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     2581 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2641 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2701 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2761 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2821 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2881 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2941 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3001 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3061 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3121 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3181 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3241 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3301 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3361 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3421 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3481 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3661 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3721 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3781 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3841 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3901 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3961 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4021 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4081 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4141 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4201 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4261 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4321 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4381 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4441 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4501 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4561 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4621 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4681 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4741 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4801 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4861 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4921 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4981 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5041 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5101 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5161 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5221 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5281 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
