LOCUS       sequence-466625-        6208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV cloning vector for Flpo-dependent expression under the CAG 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6208)
  TITLE     Ritola AAV cloning vectors
REFERENCE   2  (bases 1 to 6208)
  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"
COMMENT     Sequence Label: CAG-FRT-MCS-WPRE-SV40 - GCH3
FEATURES             Location/Qualifiers
     source          1..6208
                     /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"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1007..1147
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1018..1147
                     /label=AAV2 ITR
     polyA_signal    1253..1374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1290..1309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1344..1363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1390..1978
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1496..1507
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1905..1925
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1985..2032
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    2033..2140
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2049..2065
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2050..2066
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2100..2116)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2100..2116)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(2141..2188)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(2222..2241)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2249..3265
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2303..2325)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3266..3546)
                     /label=chicken beta-actin promoter
     enhancer        3548..3927
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   3957..4097
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3957..4086
                     /label=AAV2 ITR
     primer_bind     complement(4107..4124)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4107..4123)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4116..4138)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4336..4791
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4423..4442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4633..4654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4808..4827)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4927..4949
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4987..5005)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5073..5177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5178..5246,5247..6038)
                     /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(5396..5415)
                     /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 tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gaattgcctg caggcagctg
     1021 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     1081 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     1141 ggttcctatc gatggcgcgc ctaccacatt tgtagaggtt ttacttgctt taaaaaacct
     1201 cccacatctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1261 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1321 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1381 ctgtgtacag cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa
     1441 gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga
     1501 gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga
     1561 gcagccaagg aaaggacgat gatttccccg acaacaccac ggaattgtca gtgcccaaca
     1621 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag
     1681 ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag
     1741 tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca
     1801 actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac
     1861 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     1921 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgc
     1981 tagcgaagtt cctattccga agttcctatt ctctagaaag tataggaact tcggtaccgg
     2041 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
     2101 atccactagt tctagagcgg ccgccaccgc ggtggagctc gaagttccta tactttctag
     2161 agaataggaa cttcggaata ggaacttcac cggtcaattc tttgccaaaa tgatgagaca
     2221 gcacaataac cagcacgttg cccaggagct gtaggaaaaa gaagaaggca tgaacatggt
     2281 tagcagaggc tctagagccg ccggtcacac gccagaagcc gaaccccgcc ctgccccgtc
     2341 ccccccgaag gcagccgtcc ccccgcggac agccccgagg ctggagaggg agaaggggac
     2401 ggcggcgcgg cgacgcacga aggccctccc cgcccatttc cttcctgccg gcgccgcacc
     2461 gcttcgcccc gcgcccgcta gagggggtgc ggcggcgcct cccagatttc ggctccgcac
     2521 agatttggga caaaggaagt ccctgcgccc tctcgcacga ttaccataaa aggcaatggc
     2581 tgcggctcgc cgcgcctcga cagccgccgg cgctccgggg gccgccgcgc ccctcccccg
     2641 agccctcccc ggcccgaggc ggccccgccc cgcccggcac ccccacctgc cgccaccccc
     2701 cgcccggcac ggcgagcccc gcgccacgcc ccgtacggag ccccgcaccc gaagccgggc
     2761 cgtgctcagc aactcgggga ggggggtgca gggggggttg cagcccgacc gacgcgccca
     2821 caccccctgc tcaccccccc acgcacacac cccgcacgca gcctttgttc ccctcgcagc
     2881 cccccccgca ccgcggggca ccgcccccgg ccgcgctccc ctcgcgcaca ctgcggagcg
     2941 cacaaagccc cgcgccgcgc ccgcagcgct cacagccgcc gggcagcgcg gagccgcacg
     3001 cggcgctccc cacgcacaca cacacgcacg caccccccga gccgctcccc ccgcacaaag
     3061 ggccctcccg gagcccctca aggctttcac gcagccacag aaaagaaaca agccgtcatt
     3121 aaaccaagcg ctaattacag cccggaggag aagggccgtc ccgcccgctc acctgtggga
     3181 gtaacgcggt cagtcagagc cggggcgggc ggcgcgaggc ggcggcggag cggggcacgg
     3241 ggcgaaggca gcgcgcagcg actcccgccc gccgcgcgct tcgcttttta tagggccgcc
     3301 gccgccgccg cctcgccata aaaggaaact ttcggagcgc gccgctctga ttggctgccg
     3361 ccgcacctct ccgcctcgcc ccgccccgcc cctcgccccg ccccgccccg cctggcgcgc
     3421 gccccccccc cccccccccc gcccccatcg ctgcacaaaa taattaaaaa ataaataaat
     3481 acaaaattgg gggtggggag gggggggaga tggggagagt gaagcagaac gtggggctca
     3541 cctcgaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     3601 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     3661 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     3721 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     3781 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     3841 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     3901 attaataact agtcaataat caatgtcgag ggttaccgcc taggatcgat agatctagga
     3961 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4021 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4081 gcgcagctgc ctgcaggcag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     4141 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4201 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4261 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4321 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4381 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4441 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4501 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4561 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4621 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     4681 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4741 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4801 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4861 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4921 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4981 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5041 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5101 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5161 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5221 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5281 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5341 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5401 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5461 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5521 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5581 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5641 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5701 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5761 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5821 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5881 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5941 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6001 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6061 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6121 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6181 accccgtaga aaagatcaaa ggatcttc
//
