LOCUS       sequence-383620-        6231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For high-level expression of the AAV9 cap gene, suitable for use 
            with pCMV-Rep78/68 SSM library.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6231)
  AUTHORS   Jain NK, Ogden PJ, Church GM
  TITLE     Comprehensive mutagenesis maps the effect of all single-codon 
            mutations in the AAV2 rep gene on AAV production.
  JOURNAL   Elife. 2024 Mar 27;12:RP87730. doi: 10.7554/eLife.87730.
  PUBMED    38536879
REFERENCE   2  (bases 1 to 6231)
  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"; journalName: "Elife."; 
            date: "2024-03-27"; volume: "12:RP87730. doi"; pages: " 
            10.7554/eLife.87730"
COMMENT     Sequence Label: pCMV-AAV9cap
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             2899..2910
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    3344..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3368..3387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3422..3441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3589..3605)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3589..3605)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3602..3624)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3613..3629
                     /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)."
     promoter        complement(join(3637..3643,3644..3661,3662..3667))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3682..3703
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3820..3837)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3991..4579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4071..4090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4750..5541,5542..5610))
                     /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     5373..5392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5611..5715)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5783..5801
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5839..5861)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5961..5980
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6174..6196
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6188..6205
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6189..6205
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactcgagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcggccgca attcccgggg atcgaaagag cctgctaaag caaaaaagaa
      841 gtcaccatgg ctgccgatgg ttatcttcca gattggctcg aggacaacct tagtgaagga
      901 attcgcgagt ggtgggcttt gaaacctgga gcccctcaac ccaaggcaaa tcaacaacat
      961 caagacaacg ctcgaggtct tgtgcttccg ggttacaaat accttggacc cggcaacgga
     1021 ctcgacaagg gggagccggt caacgcagca gacgcggcgg ccctcgagca cgacaaggcc
     1081 tacgaccagc agctcaaggc cggagacaac ccgtacctca agtacaacca cgccgacgcc
     1141 gagttccagg agcggctcaa agaagatacg tcttttgggg gcaacctcgg gcgagcagtc
     1201 ttccaggcca aaaagaggct tcttgaacct cttggtctgg ttgaggaagc ggctaagacg
     1261 gctcctggaa agaagaggcc tgtagagcag tctcctcagg aaccggactc ctccgcgggt
     1321 attggcaaat cgggtgcaca gcccgctaaa aagagactca atttcggtca gactggcgac
     1381 acagagtcag tcccagaccc tcaaccaatc ggagaacctc ccgcagcccc ctcaggtgtg
     1441 ggatctctta caatggcttc aggtggtggc gcaccagtgg cagacaataa cgaaggtgcc
     1501 gatggagtgg gtagttcctc gggaaattgg cattgcgatt cccaatggct gggggacaga
     1561 gtcatcacca ccagcacccg aacctgggcc ctgcccacct acaacaatca cctctacaag
     1621 caaatctcca acagcacatc tggaggatct tcaaatgaca acgcctactt cggctacagc
     1681 accccctggg ggtattttga cttcaacaga ttccactgcc acttctcacc acgtgactgg
     1741 cagcgactca tcaacaacaa ctggggattc cggcctaagc gactcaactt caagctcttc
     1801 aacattcagg tcaaagaggt tacggacaac aatggagtca agaccatcgc caataacctt
     1861 accagcacgg tccaggtctt cacggactca gactatcagc tcccgtacgt gctcgggtcg
     1921 gctcacgagg gctgcctccc gccgttccca gcggacgttt tcatgattcc tcagtacggg
     1981 tatctgacgc ttaatgatgg aagccaggcc gtgggtcgtt cgtcctttta ctgcctggaa
     2041 tatttcccgt cgcaaatgct aagaacgggt aacaacttcc agttcagcta cgagtttgag
     2101 aacgtacctt tccatagcag ctacgctcac agccaaagcc tggaccgact aatgaatcca
     2161 ctcatcgacc aatacttgta ctatctctca aagactatta acggttctgg acagaatcaa
     2221 caaacgctaa aattcagtgt ggccggaccc agcaacatgg ctgtccaggg aagaaactac
     2281 atacctggac ccagctaccg acaacaacgt gtctcaacca ctgtgactca aaacaacaac
     2341 agcgaatttg cttggcctgg agcttcttct tgggctctca atggacgtaa tagcttgatg
     2401 aatcctggac ctgctatggc cagccacaaa gaaggagagg accgtttctt tcctttgtct
     2461 ggatctttaa tttttggcaa acaaggaact ggaagagaca acgtggatgc ggacaaagtc
     2521 atgataacca acgaagaaga aattaaaact actaacccgg tagcaacgga gtcctatgga
     2581 caagtggcca caaaccacca gagtgcccaa gcacaggcgc agaccggctg ggttcaaaac
     2641 caaggaatac ttccgggtat ggtttggcag gacagagatg tgtacctgca aggacccatt
     2701 tgggccaaaa ttcctcacac ggacggcaac tttcaccctt ctccgctgat gggagggttt
     2761 ggaatgaagc acccgcctcc tcagatcctc atcaaaaaca cacctgtacc tgcggatcct
     2821 ccaacggcct tcaacaagga caagctgaac tctttcatca cccagtattc tactggccaa
     2881 gtcagcgtgg agatcgagtg ggagctgcag aaggaaaaca gcaagcgctg gaacccggag
     2941 atccagtaca cttccaacta ttacaagtct aataatgttg aatttgctgt taatactgaa
     3001 ggtgtatata gtgaaccccg ccccattggc accagatacc tgactcgtaa tctgtaataa
     3061 taaccgggca ggccatgtct gcccgtattt cgcgtaagga aatccattat gtactattta
     3121 aaaaacacaa acttttggat gttcggttta ttctttttct tttacttttt tatcatggga
     3181 gcctacttcc cgtttttccc gatttggcta catgacatca accatatcag caaaagtgat
     3241 acgggtatta tttttgccgc tatttctctg ttctcgctat tattccaacc gctgtttggt
     3301 ctgctttctg acaaactcgg cctcgactct aggcggccgc ggggatccag acatgataag
     3361 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3421 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3481 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc
     3541 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3601 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3661 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3721 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3781 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3841 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3901 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3961 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4021 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4081 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4141 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4201 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4261 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4321 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4381 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4441 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4501 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4861 atctggcccc agtgctgcaa tgatacctcg agagccacgc tcaccggctc cagatttatc
     4921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5041 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5341 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5701 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5761 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5821 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5881 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5941 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6001 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6061 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6121 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6181 acgacgttgt aaaacgacgg ccagtgaatt cgagcttgca tgcctgcagg t
//
