LOCUS       sequence-457141-        5133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV cloning vector for Flpo-dependent expression under the GFAP 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5133)
  TITLE     Ritola AAV cloning vectors
REFERENCE   2  (bases 1 to 5133)
  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: GFAP-FRT-MCS-WPRE-SV40 - GCH33
FEATURES             Location/Qualifiers
     source          1..5133
                     /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)."
     repeat_region   2882..3022
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2882..3011
                     /label=AAV2 ITR
     primer_bind     complement(3032..3049)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3032..3048)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3041..3063)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3261..3716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3348..3367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3558..3579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3733..3752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3852..3874
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3912..3930)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3998..4102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4103..4171,4172..4963)
                     /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"
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 cggtgcgagc agcggaggtg atgcgtctcc
     2221 tctccaacct gctctggctc tgctcgctcc tgggatgcga gggctttatg aaggagtggg
     2281 ctagactggc gatgcccggg tgcccctggc aacaccccct ggcatagcct gagggggtgg
     2341 ggttgggccg cagcccagct atggggagag ctctcccctc acccatttgt gtccttggac
     2401 ttctgggaag cagacagccc tcctctcccc ccctgccacc cactgtccat gcctgggcat
     2461 cccctgatcc cctttcctgg gcacaggctg aatagagcct tgttctccac cgccggtggc
     2521 gccacacagc aagggcagcc cctagggggc tgtgctgctt ttatcccaag atgccaggct
     2581 gtcaggctgg ggtgcaatgc aagccccctg ctcaatgggc ttctcggaag gcatttgagg
     2641 ggggctgggg tgcggcggga agcaggcact gtgcccaccc cgaacattgt gtctgtgcca
     2701 aggtgagtca ttcactgggc atgaagagga ggccctgggg ggcggccagc ccaatgctgc
     2761 caggtctgct tgcagacaag gcctggctgt ctgcctcctc cccagctcac agagccagct
     2821 caggcccccg agcctctgtc agagcagcgt cccctactcc acaccaggat atgttagatc
     2881 taggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2941 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3001 cgagcgcgca gctgcctgca ggcagcttgg cactggccgt cgttttacaa cgtcgtgact
     3061 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3121 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3181 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3241 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3301 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3361 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3421 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3481 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3541 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3601 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3661 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3721 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3781 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3841 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3901 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3961 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4021 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4081 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4141 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4201 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4261 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4321 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4381 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4441 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4501 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4561 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4621 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4681 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4741 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4801 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4861 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4921 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4981 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5041 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5101 gtcagacccc gtagaaaaga tcaaaggatc ttc
//
