LOCUS       sequence-473029-        5696 bp    DNA     circular SYN 16-MAY-2025
DEFINITION  An AAV genome that ubiquitously expresses the fluorescent protein 
            cyOFP from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5696)
  TITLE     Schnitzer lab plasmids 2025
REFERENCE   2  (bases 1 to 5696)
  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-cyOFP-WPRE
FEATURES             Location/Qualifiers
     source          1..5696
                     /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..1799
                     /codon_start=1
                     /product="orange-red fluorescent protein that is excitable 
                     by cyan light and suitable as an acceptor for blue-emitting
                     luciferase (Chu et al., 2016)"
                     /label=CyOFP1
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTN
                     RIKVVEGGPLPFAFDILATHFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKVRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KALKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     misc_feature    1823..2411
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1876..1896)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2294..2305)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2443..2919
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2642..2661)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2959..3099
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2959..3088
                     /label=AAV2 ITR
     rep_origin      3174..3629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3471..3492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3646..3665)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3765..3787
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3825..3843)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3911..4015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4016..4084,4085..4876)
                     /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(4234..4253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5047..5635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5536..5555
                     /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 gtgagcaagg gcgaggagct gatcaaggag aacatgagaa
     1141 gcaagctgta cctggaaggc agcgtgaacg gccaccagtt caagtgcacc cacgaagggg
     1201 agggcaagcc ctacgagggc aagcagacca acaggatcaa ggtggtggag ggaggccccc
     1261 tgccgttcgc attcgacatc ctggccaccc actttatgta cgggagcaag gtgttcatca
     1321 agtaccccgc cgacctcccc gattatttta agcagtcctt ccctgagggc ttcacatggg
     1381 agagagtcat ggtgttcgaa gacgggggcg tgctgaccgc cacccaggac accagcctcc
     1441 aggacggcga gctcatctac aacgtcaagg tcagaggggt gaacttccca gccaacggcc
     1501 ccgtgatgca gaagaaaaca ctgggctggg agcccagcac cgagaccatg taccccgctg
     1561 acggcggcct ggaaggcaga tgcgacaagg ccctgaagct cgtgggcggg ggccacctgc
     1621 acgtcaactt caagaccaca tacaagtcca agaaacccgt gaagatgccc ggcgtccact
     1681 acgtggaccg cagactggaa agaatcaagg aggccgacaa cgagacctac gtcgagcagt
     1741 acgagcacgc tgtggccaga tactccaacc tgggcggagg catggacgag ctgtacaagt
     1801 aatgaattcg aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     1861 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1921 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1981 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2041 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2101 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2161 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2221 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     2281 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2341 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2401 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     2461 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2521 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2581 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2641 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2701 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2761 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     2821 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     2881 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     2941 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3001 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3061 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3121 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3181 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3241 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3301 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3361 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3421 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3481 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     3541 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3601 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3661 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3721 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3781 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3841 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3901 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3961 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4021 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4081 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4141 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4201 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4261 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4321 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4381 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4441 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4501 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4561 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4621 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4681 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4741 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4801 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4861 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4921 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4981 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5041 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5101 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5161 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5221 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5281 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5341 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5401 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5461 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5521 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5581 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5641 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
