LOCUS       sequence-421937-        6015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FlpO expression under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6015)
  TITLE     AAV expression plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6015)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CAG-FlpO
FEATURES             Location/Qualifiers
     source          1..6015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   826..955
                     /label=AAV2 ITR
     polyA_signal    1117..1238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1154..1173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1208..1227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1281..1869
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1387..1398
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1796..1816
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1882..3153,3154..3174,3175..3180))
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     regulatory      3177..3186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3222..3241)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3303..3325)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3604..3887)
                     /label=chicken beta-actin promoter
     repeat_region   4198..4327
                     /label=AAV2 ITR
     rep_origin      4399..4854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4486..4505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4696..4717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4880..4984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4985..5053,5054..5845)
                     /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(5203..5222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg cgctcgctcg
      841 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      901 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      961 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccttatc
     1021 gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca tctccccctg
     1081 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg
     1261 cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg
     1321 gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc
     1381 cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc
     1441 aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc
     1501 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     1561 cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat
     1621 gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac
     1681 aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat
     1741 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
     1801 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
     1861 aggttgatta tcgataagct ttcagatccg cctgttgatg tagctgctca ggtagtccag
     1921 cacctcctgg ctgatgatgc cgttccaggc ggggtatctg atgctgccct cggcgctgcc
     1981 cttcagctgc tcgatgtgct gccactcctc gatggggttg gtctcgtcct tcagggcgat
     2041 catctccttg ctgatggggt cgtaggcgta gtacctggac accagggcga agtagtggtc
     2101 ggggatggcg gtgatctggt gggtgtaggt ggtcctggcc acggcggagg ccctcttgtc
     2161 gctccagttg cccaccacgt ttgtcagctc ggtcaggccc ttcatgctca gaaagctggt
     2221 catcaggtgc ctgccgatgt ggctcttagg gccgttcttg atagcgaaga tggggtaggg
     2281 ggcgttcttc ttcagggcct tgttgtagct gcgcaccagg ttgtccttca gcagctggta
     2341 ctcctgcttg ttgctgctgc tgttgccggt cctgttcact ctcttcagca cgggctcgct
     2401 gttcctcagg aactcgtcca ggtacaccag ggggtcgatc ctgcctctgg cgctgaaaaa
     2461 gtagatgtgc ctggacacgc ttgtcttggt ctcggtcacc aggcactgaa tgatcacgcc
     2521 caggtacttg ttctgcacca gcttgaagct cttggggtcc acgttcttga tgtcgctgaa
     2581 cctgccgcag ttgatgaatg tggccaggaa caggaactgg tacagggtct tggtcttggt
     2641 gaacctgctg gtgtactcga agctgttcag gatcttctcg gtgatctccc agatgctctc
     2701 gccctcggac agcagggcct tcagcatctt cttgctgtgg ctgttgccct tgtcggcctc
     2761 ctcgctgctc tcgaactgca gctgcaggct ggacacgatg tcggtgatgt cgctctggtg
     2821 cttctggccg ttgtaaggga tgatggtgaa ctcccaggcg gggatcagct tcttcaggct
     2881 ggcctccagg atggtggcct tctgggtctt gtacttgaac tgcaggctct tgttcacgat
     2941 gtcgaagctc aggctgttgc tgatgatggt gttgtagctc atgaaggtgg ccctcttgat
     3001 ggcggtgccg ttgtgggtga tcatccagca caggtaggtc agctcggcgg cacagctggc
     3061 gatcttctcg ccgctgggcc tctcgaatct ctccacgaac tgccgcacca gcaccttggg
     3121 gggggtcttg cacaggatgt cgaactggct catcaccttc ctcttcttct taggagccat
     3181 ggtggcggcg gatccaattc tttgccaaaa tgatgagaca gcacaataac cagcacgttg
     3241 cccaggagct gtaggaaaaa gaagaaggca tgaacatggt tagcagaggc tctagagccg
     3301 ccggtcacac gccagaagcc gaaccccgcc ctgccccgtc ccccccgaag gcagccgtcc
     3361 ccccgcggac agccccgagc cgctcccccc gcacaaaggg ccctcccgga gcccctcaag
     3421 gctttcacgc agccacagaa aagaaacaag ccgtcattaa accaagcgct aattacagcc
     3481 cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg
     3541 gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac
     3601 tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa
     3661 aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc
     3721 gccccgcccc tcgccccgcc ccgccccgcc tggcgcgcgc cccccccccc cccccccccc
     3781 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga
     3841 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc
     3901 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     3961 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     4021 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     4081 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatggtac
     4141 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     4201 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4261 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4321 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4381 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4441 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4501 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4561 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4621 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4681 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4741 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4801 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4861 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4981 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5041 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5101 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5161 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5221 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5281 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5341 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5401 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5461 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5521 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5581 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5641 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5701 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5761 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5821 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5881 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5941 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6001 gatcaaagga tcttc
//
