LOCUS       sequence-473030-        5709 bp    DNA     circular SYN 16-MAY-2025
DEFINITION  An AAV genome that ubiquitously expresses the fluorescent protein 
            mRuby2 from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5709)
  TITLE     Schnitzer lab plasmids 2025
REFERENCE   2  (bases 1 to 5709)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV-CAG-mRuby2-WPRE
FEATURES             Location/Qualifiers
     source          1..5709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        404..681
                     /label=chicken beta-actin promoter
     primer_bind     961..983
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1045..1064
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1098..1808
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    1836..2424
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1889..1909)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2307..2318)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2456..2932
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2655..2674)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2972..3112
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2972..3101
                     /label=AAV2 ITR
     rep_origin      3187..3642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3274..3293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3484..3505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3659..3678)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3778..3800
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3838..3856)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3924..4028
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4029..4097,4098..4889)
                     /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(4247..4266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5060..5648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5549..5568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggacta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
      421 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      481 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg
      541 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
      601 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
      661 aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg tgccccgctc
      721 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      781 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
      841 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
      901 gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
      961 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     1021 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     1081 gcaaagaatt ggatccgatg gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta
     1141 tgaaggtggt catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag
     1201 aaggcaatcc gtacatggga actcaaacca tgaggatcaa agtcatcgag ggaggacccc
     1261 tgccatttgc ctttgacatt cttgccacgt cgttcatgta tggcagccgt acttttatca
     1321 agtacccgaa aggcattcct gatttcttta aacagtcctt tcctgagggt tttacttggg
     1381 aaagagttac gagatacgaa gatggtggag tcgtcaccgt catgcaggac accagccttg
     1441 aggatggctg tctcgtttac cacgtccaag tcagaggggt aaactttccc tccaatggtc
     1501 ccgtgatgca gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag
     1561 atggtggtct gaggggatac actcatatgg cactgaaagt tgatggtggt ggccatctgt
     1621 cttgctcttt cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg
     1681 gtatccatgc cgttgatcac cgcctggaaa ggttagagga aagtgacaat gaaatgttcg
     1741 tagtacaacg cgaacacgca gttgccaagt tcgccgggct tggtggtggg atggacgagc
     1801 tgtacaagta agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     1861 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     1921 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     1981 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2041 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2101 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2161 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2221 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     2281 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2341 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2401 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     2461 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     2521 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     2581 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     2641 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     2701 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     2761 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     2821 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     2881 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     2941 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3001 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3061 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3121 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3181 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3241 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3301 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3361 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3421 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3481 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     3541 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3601 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3661 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3721 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3781 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3841 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3901 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3961 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4021 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4081 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4141 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4201 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4261 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4321 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4381 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4441 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4501 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4561 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4621 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4681 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4741 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4801 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4861 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4921 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4981 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5041 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5101 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5161 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5221 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5281 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5341 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5401 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5461 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5521 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5581 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5641 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5701 ctcacatgt
//
