LOCUS       sequence-330202-        6111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV, encodes monomeric ultrastable GFP (muGFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6111)
  AUTHORS   Scott DJ, Gunn NJ, Yong KJ, Wimmer VC, Veldhuis NA, Challis LM, 
            Haidar M, Petrou S, Bathgate RAD, Griffin MDW
  TITLE     A Novel Ultra-Stable, Monomeric Green Fluorescent Protein For Direct
            Volumetric Imaging of Whole Organs Using CLARITY.
  JOURNAL   Sci Rep. 2018 Jan 12;8(1):667. doi: 10.1038/s41598-017-18045-y.
  PUBMED    29330459
REFERENCE   2  (bases 1 to 6111)
  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: "Sci Rep."; 
            date: "2018-01-12"; volume: "8(1):667. doi"; pages: " 
            10.1038/s41598-017-18045-y"
COMMENT     Sequence Label: pAM-DCA-muGFP
FEATURES             Location/Qualifiers
     source          1..6111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     promoter        466..743
                     /label=chicken beta-actin promoter
     primer_bind     1022..1044
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1106..1125
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1205..1918
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVLCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     YFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     DVTAAGITHGMDELYK"
     primer_bind     complement(1235..1263)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     misc_feature    1937..2525
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1990..2010)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2408..2419)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2552..2569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2558..2782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(2992..3321)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3006..3141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3306..3326
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3567..4155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3647..3666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4326..5117,5118..5186))
                     /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     4949..4968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5187..5291)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5359..5377
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5415..5437)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5537..5556
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5770..5789
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6079..6097
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6079..6096
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accatcctac tactagtacg tcgtaccatt gacgtcaata atgacgtatg
      241 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      301 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      361 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      421 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      481 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      541 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccrgggg
      601 gggggggggg gggsgggggg gggggsgggg sgrggcggag aggtgcggcg gcagccaatc
      661 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      721 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      781 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      841 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      901 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
      961 ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg
     1021 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc
     1081 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
     1141 ggcaaagaat tggatcttcg atcgaggaat tcaagctgct agcaaggatc caccggtcgc
     1201 caccatgagc aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
     1261 tggtgatgtt aatgggcaca aattttctgt ccgtggagag ggtgaaggtg atgctacaaa
     1321 cggaaaactc acccttaaat ttatttgcac tactggaaaa ctacctgttc cgtggccaac
     1381 acttgtcact actctgacct atggtgttct atgcttttcc cgttatccgg atcacatgaa
     1441 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc
     1501 tttcaaagat gacgggacct acaagacgcg tgctgaagtc aagtttgaag gtgataccct
     1561 tgttaatcgt atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttctcggaca
     1621 caaactggag tacaacttta actcacacaa tgtatacatc acggcagaca aacaaaagaa
     1681 tggaatcaaa gcgtacttca aaattcgcca caacgttgaa gatggttccg ttcaactagc
     1741 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     1801 ttacctctcg acacaatctg ttctttcgaa agatcccaac gaaaagcgtg accacatggt
     1861 ccttcttgag gatgtaactg ctgctgggat tacacatggc atggatgaac tctacaaata
     1921 atagaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1981 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2041 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2101 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2161 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2221 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2281 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2341 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     2401 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2461 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2521 cccgcatcga taccgtcgac tcgctgatca gcctcgactg tgccttctag ttgccagcca
     2581 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2641 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2701 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2761 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgactaga
     2821 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     2881 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     2941 ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagagct ttttgcaaaa
     3001 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     3061 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     3121 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     3181 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     3241 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     3301 tccacaccct aactgacaca cattccacag ctgcattaat gaatcggcca acgcgcgggg
     3361 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3421 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3481 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3541 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3601 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3661 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3721 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3781 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3841 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3901 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3961 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4021 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4081 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4141 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4201 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4261 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4321 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4381 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4441 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4501 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4561 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4621 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4681 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4741 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4801 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4861 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4921 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4981 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5041 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5101 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5161 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5221 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5281 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5341 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5401 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5461 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5521 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca ttcgacgctc
     5581 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5641 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5701 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5761 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5821 gccggccacg atgcgtccgg cgtagaggat ctggctagcg atgaccctgc tgattggttc
     5881 gctgaccatt tccgggtgcg ggacggcgtt accagaaact cagaaggttc gtccaaccaa
     5941 accgactctg acggcagttt acgagagaga tgatagggtc tgcttcagta agccagatgc
     6001 tacacaatta ggcttgtaca tattgtcgtt agaacgcggc tacaattaat acataacctt
     6061 atgtatcata cacatacgat ttaggtgaca ctatagaata cacggaatta a
//
