LOCUS       sequence-459148-        6110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV cloning vector for expression under the CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6110)
  TITLE     Ritola AAV cloning vectors
REFERENCE   2  (bases 1 to 6110)
  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-MCSrev-WPRE-SV40 - GCH4
FEATURES             Location/Qualifiers
     source          1..6110
                     /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"
     misc_feature    1985..2092
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2009..2025
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2009..2025
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2059..2075)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2060..2076)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2126..2145)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2153..3169
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2207..2229)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3170..3448)
                     /label=chicken beta-actin promoter
     enhancer        3450..3829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   3859..3999
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3859..3988
                     /label=AAV2 ITR
     primer_bind     complement(4009..4026)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4009..4025)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4018..4040)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4238..4693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4325..4344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4535..4556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4710..4729)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4829..4851
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4889..4907)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4975..5079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5080..5148,5149..5940)
                     /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(5298..5317)
                     /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 ggttgattac
     1981 cggtgagctc caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga
     2041 attcgatatc aagcttatcg ataccgtcga cctcgagggg gggcccggta ccgctagtca
     2101 attctttgcc aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga
     2161 aaaagaagaa ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga
     2221 agccgaaccc cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc
     2281 gaggctggag agggagaagg ggacggcggc gcggcgacgc acgaaggccc tccccgccca
     2341 tttccttcct gccggcgccg caccgcttcg ccccgcgccc gctagagggg gtgcggcggc
     2401 gcctcccaga tttcggctcc gcacagattt gggacaaagg aagtccctgc gccctctcgc
     2461 acgattacca taaaaggcaa tggctgcggc tcgccgcgcc tcgacagccg ccggcgctcc
     2521 gggggccgcc gcgccccycc cccgagcccy ccccggcccg mggcggcccc gccccgcccg
     2581 gcacccccac ctgccgccac cccccgcccg gcacggcgag ccccgcgcca cgccccgtac
     2641 ggagccccgc acccgaagcc gggccgtgct cagcaactcg gggagggggg tgcagggggg
     2701 gttgcagccc gaccgacgcg cccacacccc ctgctcaccc ccccacgcac acaccccgca
     2761 cgcagccttt gttcccctcg cagccccccc cgcaccgcgg ggcaccgccc ccggccgcgc
     2821 tcccctcgcg cacactgcgg agcgcacaaa gccccgcgcc gcgcccgcag cgctcacagc
     2881 cgccgggcag cgcggagccg cacgcggcgc tccccacgca cacacacacg cacgcacccc
     2941 ccgagccgct ccccccgcac aaagggccct cccggagccc ctcaaggctt tcacgcagcc
     3001 acagaaaaga aacaagccgt cattaaacca agcgctaatt acagcccgga ggagaagggc
     3061 cgtcccgccc gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg
     3121 aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg
     3181 cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga
     3241 gcgcgccgct ctgattggct gccgccgcac ctctccscct csccccsccc cccccctccc
     3301 cccccccccc cccccctsgc gcgcgccccc cccccccccc ccgcccccat cgctgcacaa
     3361 aataattaaa aaataaataa atacaaaatt gggggtgggg agggggggga gatggggaga
     3421 gtgaagcaga acgtggggct cacctcgacc atggtaatag cgatgactaa tacgtagatg
     3481 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     3541 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     3601 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     3661 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     3721 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     3781 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtcg agggttaccg
     3841 cctaggatcg atagatctag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3901 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3961 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggc agcttggcac tggccgtcgt
     4021 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     4081 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4141 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     4201 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4261 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4321 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4381 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4441 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4501 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4561 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     4621 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4681 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4741 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4801 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4861 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4921 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4981 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5041 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5101 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5161 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5221 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5281 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5341 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5401 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5461 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5521 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5581 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5641 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5701 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5761 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5821 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5881 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5941 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6001 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6061 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
