LOCUS       sequence-447425-        4893 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 4893)
  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 4893)
  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: HCT14_rAAV-eHGT_1051m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..4893
                     /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      905..914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(911..913,914..916,917..1627)
                     /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(956..977)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1217..1236)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1564..1585
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1720..1740)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1927..2038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2156..2296
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2156..2285
                     /label=AAV2 ITR
     rep_origin      2371..2826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2458..2477)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2668..2689
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2843..2862)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2962..2984
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3022..3040)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3108..3212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3213..3281,3282..4073)
                     /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(3431..3450)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4244..4832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4733..4752
                     /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 gccggcgctg attttcccaa
      181 gggcagttta cagagcagcc ctcccctgtg caccactgca ggagacttag tattaagttg
      241 cctgccctgg ttctgactgg atgacattgt tccaaaaagg atgcccccaa gatcttccca
      301 ctagaaactt gcacagggca cgttttaggg tgtgcttgcc gccatcctgc cctgtatgag
      361 caggacctaa cagggttaat tagggccttg gctatagatt tggagctcag caaagctctg
      421 gggtcctcaa gcccagctgg catgggggta ggaatggctg gcatctcctg ccgggttggg
      481 tgaaggccat agcagtatct ggctgcccag agcgttctgt atttgtaaaa cacagtgttc
      541 caacactggg ccaggctgga tatagccagg ccttgtccct atctggcctg gattgtccta
      601 tcaccttgtg ggggtaaagc tggcagaagg agcagttggg gcattgtccc aaggagatat
      661 taagcggcca tgaggaagac acaggccctg gaacagacag cgggtcactc tccagttcag
      721 tgctcgcctc tgtgctcctg actgtgtacc cttactgtgt ggaatgcgga tgctgatccc
      781 ccatggttgt tccacatttg acccgccggc ggagctcggg ctgggcataa aagtcagggc
      841 agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc
      901 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      961 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1021 tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc
     1081 ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga
     1141 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1201 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1261 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1321 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa
     1381 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
     1441 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1501 cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga
     1561 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1621 gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga
     1681 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1741 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     1801 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     1861 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     1921 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     1981 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2041 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2101 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     2161 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2221 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2281 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2341 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     2401 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     2461 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2521 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2581 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2641 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2701 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     2761 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     2821 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     2881 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2941 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3001 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3061 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3121 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3181 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3241 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3301 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3361 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3421 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3481 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3541 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3601 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3661 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3721 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3781 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3841 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3901 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3961 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4021 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4081 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4141 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4201 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4261 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4321 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4381 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4441 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4501 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4561 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4621 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4681 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4741 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4801 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4861 tcctggcctt ttgctggcct tttgctcaca tgt
//
