LOCUS       sequence-479178-        5893 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  AAV cloning vector for Flp dependent expression under the CamKII 
            promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  TITLE     Ritola Lab plasmids
REFERENCE   2  (bases 1 to 5893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 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 - GCH21.1
FEATURES             Location/Qualifiers
     source          1..5893
                     /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    2061..2108
                     /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    2109..2216
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2125..2141
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2126..2142
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2176..2192)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2176..2192)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(2293..2340)
                     /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(2347..3635)
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     repeat_region   3642..3782
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3642..3771
                     /label=AAV2 ITR
     primer_bind     complement(3792..3809)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3792..3808)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3801..3823)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4021..4476
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4108..4127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4318..4339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4493..4512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4612..4634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4672..4690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4758..4862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4863..4931,4932..5723)
                     /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(5081..5100)
                     /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 cttcaaatag tataggaact tcttggatca
     2041 aaataggagc gatcgcaatg gaagttccta ttccgaagtt cctattctct agaaagtata
     2101 ggaacttcgg taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat
     2161 tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg gagctcgaag
     2221 ttcctatact atttgaagaa taggaacttc ggaataggaa cttcagagcg ctttggccgc
     2281 tccggccaaa gggaagttcc tatactttct agagaatagg aacttcggaa taggaacttc
     2341 accggtgctg cccccagaac taggggccac tcgcctgccc gtgctcctga gtgcaaacgg
     2401 agaaccggct tgactgacga gcttggggct tctgagcagg gcactgtggc tgctcacagc
     2461 ctcgcgagcc ttcgtcagca tccaggtccc catggcaacc acctccgaaa cgccctgttc
     2521 ccgttgcccc ggtaactgcc tccccaacac ctgcctgcct ttcactttaa aacctgctcc
     2581 tctttgccct ggccttatat acacaatact gcaggactgt gtgggcccag gagcaagtgg
     2641 accctgttcc cccagggtga tgtactgagg gggttggaga aggggatgcg aacaaactta
     2701 gtccacaagt gatcatcgct cttcctacct cccccacctc caactatagg gcccttttaa
     2761 cagacgggat gggattaagt ggggcaaaga ggcagttgct atggtaacgg ctagtgggtg
     2821 ccacattctg ggatggtcct tgagaaagac aggatgtagt taccctggca acttcatctc
     2881 ctctgaagag acaggtattg ccccgttgcc ctggcaacag ctgggttcta aggtttacag
     2941 gggaagctaa ttcatctctc cttcccttcc cccttgtagt ctctgtaatg gtttctttgc
     3001 ttcctctggg aattggtctg gttgacttgg cctctgtctc tgtccttgtt tctttccagc
     3061 tctccctgct ttttagccat tgtgcatatc acaccgccac cttagcgtga gaagaagtac
     3121 caaacagacc cagaatgctg acggaatgac tctgtctgct ttctcacccc tatctgtcac
     3181 cactgagcag cagggcccct cctgaggcct ctgccacagc tctccccacc catcctccca
     3241 gagggaccat gaagatttgg tctcctgggg ctcctgtccc agtttctgct tgtgcttccc
     3301 gtcaaggtag gactggagag aggcaactat aatgaggtga acgtggctgg actcacccag
     3361 ggctgggcgg gtggtgtaga tgtaccctgc cctggactct tctttcctaa actcattggt
     3421 catctgccct aatgagttca ctccttctgt atatcgtact tgcatgcccc caaggcatgg
     3481 aatttaagat tagtaatctg agttcaagtt ctagctctga catttctgat gtgaccctgg
     3541 gcaggtcact gaggaagctc tctgcacccc catctcattt tctagaaaat cagaagaaga
     3601 accccatgga gatgaagtga cctaaggcca taatgagatc taggaacccc tagtgatgga
     3661 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3721 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3781 ggcagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3841 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3901 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     3961 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4021 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4081 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4141 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4201 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4261 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4321 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     4381 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4441 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4501 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4561 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4621 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4681 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4741 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4801 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4861 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4921 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4981 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5041 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5101 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5161 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5221 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5281 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5341 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5401 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5461 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5521 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5581 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5641 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5701 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5761 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5821 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5881 tcaaaggatc ttc
//
