LOCUS       sequence-457144-        5741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV cloning vector for Flpo-dependent expression under the CaMKII 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5741)
  TITLE     Ritola AAV cloning vectors
REFERENCE   2  (bases 1 to 5741)
  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: CaMKII-FRT-MCS-WPRE-SV40 - GCH21
FEATURES             Location/Qualifiers
     source          1..5741
                     /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)."
     promoter        complement(2195..3483)
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     repeat_region   3490..3630
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3490..3619
                     /label=AAV2 ITR
     primer_bind     complement(3640..3657)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3640..3656)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3649..3671)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3869..4324
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3956..3975)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4166..4187
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4341..4360)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4460..4482
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4520..4538)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4606..4710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4711..4779,4780..5571)
                     /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(4929..4948)
                     /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 cggtgctgcc cccagaacta ggggccactc
     2221 gcctgcccgt gctcctgagt gcaaacggag aaccggcttg actgacgagc ttggggcttc
     2281 tgagcagggc actgtggctg ctcacagcct cgcgagcctt cgtcagcatc caggtcccca
     2341 tggcaaccac ctccgaaacg ccctgttccc gttgccccgg taactgcctc cccaacacct
     2401 gcctgccttt cactttaaaa cctgctcctc tttgccctgg ccttatatac acaatactgc
     2461 aggactgtgt gggcccagga gcaagtggac cctgttcccc cagggtgatg tactgagggg
     2521 gttggagaag gggatgcgaa caaacttagt ccacaagtga tcatcgctct tcctacctcc
     2581 cccacctcca actatagggc ccttttaaca gacgggatgg gattaagtgg ggcaaagagg
     2641 cagttgctat ggtaacggct agtgggtgcc acattctggg atggtccttg agaaagacag
     2701 gatgtagtta ccctggcaac ttcatctcct ctgaagagac aggtattgcc ccgttgccct
     2761 ggcaacagct gggttctaag gtttacaggg gaagctaatt catctctcct tcccttcccc
     2821 cttgtagtct ctgtaatggt ttctttgctt cctctgggaa ttggtctggt tgacttggcc
     2881 tctgtctctg tccttgtttc tttccagctc tccctgcttt ttagccattg tgcatatcac
     2941 accgccacct tagcgtgaga agaagtacca aacagaccca gaatgctgac ggaatgactc
     3001 tgtctgcttt ctcaccccta tctgtcacca ctgagcagca gggcccctcc tgaggcctct
     3061 gccacagctc tccccaccca tcctcccaga gggaccatga agatttggtc tcctggggct
     3121 cctgtcccag tttctgcttg tgcttcccgt caaggtagga ctggagagag gcaactataa
     3181 tgaggtgaac gtggctggac tcacccaggg ctgggcgggt ggtgtagatg taccctgccc
     3241 tggactcttc tttcctaaac tcattggtca tctgccctaa tgagttcact ccttctgtat
     3301 atcgtacttg catgccccca aggcatggaa tttaagatta gtaatctgag ttcaagttct
     3361 agctctgaca tttctgatgt gaccctgggc aggtcactga ggaagctctc tgcaccccca
     3421 tctcattttc tagaaaatca gaagaagaac cccatggaga tgaagtgacc taaggccata
     3481 atgagatcta ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3541 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3601 tgagcgagcg agcgcgcagc tgcctgcagg cagcttggca ctggccgtcg ttttacaacg
     3661 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3721 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3781 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3841 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3901 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3961 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4021 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4081 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4141 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4201 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4261 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4321 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4381 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4441 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4501 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4561 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4621 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4681 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4741 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4801 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4861 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4921 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4981 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5041 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5101 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5161 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5221 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5281 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5341 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5401 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5461 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5521 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5581 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5641 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5701 cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
