LOCUS       sequence-390030-        6228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For high-level expression of the AAV2 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 6228)
  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 6228)
  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-AAV2cap
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    248..382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(272..291)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     326..345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(493..509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(493..509)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(506..528)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    517..533
                     /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(541..547,548..565,566..571))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    586..607
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(724..741)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(895..1483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(975..994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1654..2445,2446..2514))
                     /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     2277..2296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2515..2619)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2687..2705
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2743..2765)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2865..2884
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3078..3100
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3092..3109
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3093..3109
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3136..3439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3440..3643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3594..3614
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3640..3664
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3781..3877
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             6031..6042
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
ORIGIN
        1 aggaaatcca ttatgtacta tttaaaaaac acaaactttt ggatgttcgg tttattcttt
       61 ttcttttact tttttatcat gggagcctac ttcccgtttt tcccgatttg gctacatgac
      121 atcaaccata tcagcaaaag tgatacgggt attatttttg ccgctatttc tctgttctcg
      181 ctattattcc aaccgctgtt tggtctgctt tctgacaaac tcggcctcga ctctaggcgg
      241 ccgcggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      301 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      361 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      421 gggggaggtg tgggaggttt tttcggatcc tctagagtcg acctgcaggc atgcaagctt
      481 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
      541 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
      601 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
      661 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
      721 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      781 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      841 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      901 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      961 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1021 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1081 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1141 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1201 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1261 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1321 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1381 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1441 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1501 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1561 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1621 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1681 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1741 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     1801 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1861 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1921 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1981 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2041 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2101 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2161 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2221 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2281 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2341 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2401 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2461 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2521 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2581 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     2641 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     2701 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     2761 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2821 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     2881 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     2941 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     3001 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     3061 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
     3121 tgcatgcctg caggtcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     3181 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     3241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     3301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     3361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     3421 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     3481 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     3541 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     3601 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     3661 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     3721 cagcctccgg actctagagg atccggtact cgaggaactg aaaaaccaga aagttaactg
     3781 gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg tgcaaatcaa
     3841 agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag tgttacttct
     3901 gctctaaaag ctgcggaatt gtacccgcgg ccgcaattcc cggggatcga aagagcctgc
     3961 taaagcaaaa aagaagtcac catggctgcc gatggttatc ttccagattg gctcgaggac
     4021 actctctctg aaggaataag acagtggtgg aagctcaaac ctggcccacc accaccaaag
     4081 cccgcagagc ggcataagga cgacagcagg ggtcttgtgc ttcctgggta caagtacctc
     4141 ggacccttca acggactcga caagggagag ccggtcaacg aggcagacgc cgcggccctc
     4201 gagcacgaca aagcctacga ccggcagctc gacagcggag acaacccgta cctcaagtac
     4261 aaccacgccg acgcggagtt tcaggagcgc cttaaagaag atacgtcttt tgggggcaac
     4321 ctcggacgag cagtcttcca ggcgaaaaag agggttcttg aacctctggg cctggttgag
     4381 gaacctgtta agacggctcc gggaaaaaag aggccggtag agcactctcc tgtggagcca
     4441 gactcctcct cgggaaccgg aaaggcgggc cagcagcctg caagaaaaag attgaatttt
     4501 ggtcagactg gagacgcaga ctcagtacct gacccccagc ctctcggaca gccaccagca
     4561 gccccctctg gtctgggaac taatacgatg gctacaggca gtggcgcacc aatggcagac
     4621 aataacgagg gcgccgacgg agtgggtaat tcctcgggaa attggcattg cgattccaca
     4681 tggatgggcg acagagtcat caccaccagc acccgaacct gggccctgcc cacctacaac
     4741 aaccacctct acaaacaaat ttccagccaa tcaggagcct cgaacgacaa tcactacttt
     4801 ggctacagca ccccttgggg gtattttgac ttcaacagat tccactgcca cttttcacca
     4861 cgtgactggc aaagactcat caacaacaac tggggattcc gacccaagag actcaacttc
     4921 aagctcttta acattcaagt caaagaggtc acgcagaatg acggtacgac gacgattgcc
     4981 aataacctta ccagcacggt tcaggtgttt actgactcgg agtaccagct cccgtacgtc
     5041 ctcggctcgg cgcatcaagg atgcctcccg ccgttcccag cagacgtctt catggtgcca
     5101 cagtatggat acctcaccct gaacaacggg agtcaggcag taggacgctc ttcattttac
     5161 tgcctggagt actttccttc tcagatgctg cgtaccggaa acaactttac cttcagctac
     5221 acttttgagg acgttccttt ccacagcagc tacgctcaca gccagagtct ggaccgtctc
     5281 atgaatcctc tcatcgacca gtacctgtat tacttgagca gaacaaacac tccaagtgga
     5341 accaccacgc agtcaaggct tcagttttct caggccggag cgagtgacat tcgggaccag
     5401 tctaggaact ggcttcctgg accctgttac cgccagcagc gagtatcaaa gacatctgcg
     5461 gataacaaca acagtgaata ctcgtggact ggagctacca agtaccacct caatggcaga
     5521 gactctctgg tgaatccggg cccggccatg gcaagccaca aggacgatga agaaaagttt
     5581 tttcctcaga gcggggttct catctttggg aagcaaggct cagagaaaac aaatgtggac
     5641 attgaaaagg tcatgattac agacgaagag gaaatcagga caaccaatcc cgtggctacg
     5701 gagcagtatg gttctgtatc taccaacctc cagagaggca acagacaagc agctaccgca
     5761 gatgtcaaca cacaaggcgt tcttccaggc atggtctggc aggacagaga tgtgtacctt
     5821 caggggccca tctgggcaaa gattccacac acggacggac attttcaccc ctctcccctc
     5881 atgggtggat tcggacttaa acaccctcct ccacagattc tcatcaagaa caccccggta
     5941 cctgcgaatc cttcgaccac cttcagtgcg gcaaagtttg cttccttcat cacacagtac
     6001 tccacgggac aggtcagcgt ggagatcgag tgggagctgc agaaggaaaa cagcaaacgc
     6061 tggaatcccg aaattcagta cacttccaac tacaacaagt ctgttaatgt ggactttact
     6121 gtggacacta atggcgtgta ttcagagcct cgccccattg gcaccagata cctgactcgt
     6181 aatctgtaat aataaccggg caggccatgt ctgcccgtat ttcgcgta
//
