LOCUS       sequence-466629-        6112 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 6112)
  TITLE     Ritola AAV cloning vectors
REFERENCE   2  (bases 1 to 6112)
  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-MCS-WPRE-SV40 - GCH1
FEATURES             Location/Qualifiers
     source          1..6112
                     /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"
     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     2001..2017
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2002..2018
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2052..2068)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2052..2068)
                     /label=pBluescriptSK
                     /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..3450)
                     /label=chicken beta-actin promoter
     enhancer        3452..3831
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   3861..4001
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3861..3990
                     /label=AAV2 ITR
     primer_bind     complement(4011..4028)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4011..4027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4020..4042)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4240..4695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4327..4346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4537..4558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4712..4731)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4831..4853
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4891..4909)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4977..5081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5082..5150,5151..5942)
                     /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(5300..5319)
                     /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 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     1081 cgggcggcct 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 tagcggtacc gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct
     2041 gcagcccggg ggatccacta gttctagagc ggccgccacc gcggtggagc tcaccggtca
     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 gcgcccctcc cccgagccct ccccggcccg aggcggcccc 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 ctctccgcct cgccccgccc cgcccctcgc
     3301 cccgccccgc cccgcctggc gcgcgccccc cccccccccc ccccgccccc atcgctgcac
     3361 aaaataatta aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg gagatgggga
     3421 gagtgaagca gaacgtgggg ctcacctcga ccatggtaat agcgatgact aatacgtaga
     3481 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     3541 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     3601 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     3661 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     3721 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     3781 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt cgagggttac
     3841 cgcctaggat cgatagatct aggaacccct agtgatggag ttggccactc cctctctgcg
     3901 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3961 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gcagcttggc actggccgtc
     4021 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4081 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4141 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     4201 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4261 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4321 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4381 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4441 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4501 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4561 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4621 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4681 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4741 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4801 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4861 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4921 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4981 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5041 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5101 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5161 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5221 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5281 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5341 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5401 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5461 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5521 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5581 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5641 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5701 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5761 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5821 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5881 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5941 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6001 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6061 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
