LOCUS       sequence-447433-        5042 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 5042)
  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 5042)
  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: HCT8_rAAV-eHGT_1143m-minBglobin-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5042
                     /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      1054..1063
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1060..1062,1063..1065,1066..1776)
                     /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(1105..1126)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1366..1385)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1713..1734
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1869..1889)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2076..2187
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2305..2445
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2305..2434
                     /label=AAV2 ITR
     rep_origin      2520..2975
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2607..2626)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2817..2838
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2992..3011)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3111..3133
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3171..3189)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3257..3361
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3362..3430,3431..4222)
                     /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(3580..3599)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4393..4981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4882..4901
                     /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 gccggcgcag ggattggtcc
      181 actctataca agagaaatct ttcagaaagc cgtcaccacc tttgcatctg tcatcttctg
      241 attttcaggc cctcacagcc ttagctagta acgaaccaca gcttttgtct gaggaaacct
      301 ctggtcctga ttcagagttc cagacaagtt gcttctgaca caataacaca gtcattctgc
      361 catgagaagc catttactca tgagttcaag aagccacttg cacgatgctc ttatggggtc
      421 aggttaaaca gggatggggg gttgcttttc tgaaaaaaac atcctcatca agaaaaacac
      481 tgggggtagc gatcctacat cctgccgtag ctgctgcttg ccaaagcaga cagcttttgc
      541 tattaggatg cccagccaaa cacaaatcat ccactgccac aatgagctcc ttgggacagc
      601 aggagtgttt ggaagtgaat ggctctttca gctggaaatc agcccaggtt catttccaac
      661 ccaagtcaaa atgacaaaga gtgaatttcc tgggccactc accccaaggt gtaggaacac
      721 taggtcggca gctgttatgt ttagctagtt atttaaggct ttccagcaga cggttacagt
      781 tgagcatgta accaagtcgt gcccatgccc ctcggtgaaa caatgcttct gtatcatgga
      841 cagagcctca agagacaaag ggtggggcga gggagcaagg aggaaaatca caggtcaggc
      901 acatacttat cagctgaaat gattttaatg tagtcagctt taaaatattg cacgccggcg
      961 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
     1021 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     1081 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1141 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1201 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1261 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1321 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1381 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1441 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1501 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1561 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1621 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1681 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1741 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1801 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1861 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1921 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     1981 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2041 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     2101 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2161 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2221 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     2281 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2341 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2401 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     2461 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     2521 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2581 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2641 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2701 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2761 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2821 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2881 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2941 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3001 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3061 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3121 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3181 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3241 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3301 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3361 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3421 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3481 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3541 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3601 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3661 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3721 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3781 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3841 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3901 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3961 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4021 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4081 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4141 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4201 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4261 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4321 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4381 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4441 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4501 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4561 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4621 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4681 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4741 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4801 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4861 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4921 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4981 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5041 gt
//
