LOCUS       sequence-383619-        6195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For high-level expression of the AAV5 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 6195)
  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 6195)
  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-AAV5cap
FEATURES             Location/Qualifiers
     source          1..6195
                     /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"
     polyA_signal    3308..3442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3332..3351)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3386..3405
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3553..3569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3553..3569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3566..3588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3577..3593
                     /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(3601..3607,3608..3625,3626..3631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3646..3667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3784..3801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3955..4543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4035..4054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4714..5505,5506..5574))
                     /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     5337..5356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5575..5679)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5747..5765
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5803..5825)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5925..5944
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6138..6160
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6152..6169
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6153..6169
                     /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 gtcaccatgt cttttgttga tcaccctcca gattggttgg aagaagttgg tgaaggtctt
      901 cgcgagtttt tgggccttga agcgggccca ccgaaaccaa aacccaatca gcagcatcaa
      961 gatcaagccc gtggtcttgt gctgcctggt tataactatc tcggacccgg aaacgggctc
     1021 gatcgaggag agcctgtcaa cagggcagac gaggtcgcgc gagagcacga catctcgtac
     1081 aacgagcagc ttgaggcggg agacaacccc tacctcaagt acaaccacgc ggacgccgag
     1141 tttcaggaga agctcgccga cgacacatcc ttcgggggaa acctcggaaa ggcagtcttt
     1201 caggccaaga aaagggttct cgaacctttt ggcctggttg aagagggtgc taagacggcc
     1261 cctaccggaa agcggataga cgaccacttt ccaaaaagaa agaaggctcg gaccgaagag
     1321 gactccaagc cttccacctc gtcagacgcc gaagctggac ccagcggatc ccagcagctg
     1381 caaatcccag cccaaccagc ctcaagtttg ggagctgata caatgtctgc gggaggtggc
     1441 ggcccattgg gcgacaataa ccaaggtgcc gatggagtgg gcaatgcctc gggagattgg
     1501 cattgcgatt ccacgtggat gggggacaga gtcgtcacca agtccacccg aacctgggtg
     1561 ctgcccagct acaacaacca ccagtaccga gagatcaaaa gcggctccgt cgacggaagc
     1621 aacgccaacg cctactttgg atacagcacc ccctgggggt actttgactt taaccgcttc
     1681 cacagccact ggagcccccg agactggcaa agactcatca acaactactg gggcttcaga
     1741 ccccggtccc tcagagtcaa aatcttcaac attcaagtca aagaggtcac ggtgcaggac
     1801 tccaccacca ccatcgccaa caacctcacc tccaccgtcc aagtgtttac ggacgacgac
     1861 taccagctgc cctacgtcgt cggcaacggg accgagggat gcctgccggc cttccctccg
     1921 caggtcttta cgctgccgca gtacggttac gcgacgctga accgcgacaa cacagaaaat
     1981 cccaccgaga ggagcagctt cttctgccta gagtactttc ccagcaagat gctgagaacg
     2041 ggcaacaact ttgagtttac ctacaacttt gaggaggtgc ccttccactc cagcttcgct
     2101 cccagtcaga acctgttcaa gctggccaac ccgctggtgg accagtactt gtaccgcttc
     2161 gtgagcacaa ataacactgg cggagtccag ttcaacaaga acctggccgg gagatacgcc
     2221 aacacctaca aaaactggtt cccggggccc atgggccgaa cccagggctg gaacctgggc
     2281 tccggggtca accgcgccag tgtcagcgcc ttcgccacga ccaataggat ggagctcgag
     2341 ggcgcgagtt accaggtgcc cccgcagccg aacggcatga ccaacaacct ccagggcagc
     2401 aacacctatg ccctggagaa cactatgatc ttcaacagcc agccggcgaa cccgggcacc
     2461 accgccacgt acctcgaggg caacatgctc atcaccagcg agagcgagac gcagccggtg
     2521 aaccgcgtgg cgtacaacgt cggcgggcag atggccacca acaaccagag ctccaccact
     2581 gcccccgcga ccggcacgta caacctccag gaaatcgtgc ccggcagcgt gtggatggag
     2641 agggacgtgt acctccaagg acccatctgg gccaagatcc cagagacggg ggcgcacttt
     2701 cacccctctc cggccatggg cggattcgga ctcaaacacc caccgcccat gatgctcatc
     2761 aagaacacgc ctgtgcccgg aaatatcacc agcttctcgg acgtgcccgt cagcagcttc
     2821 atcacccagt acagcaccgg gcaggtcacc gtggagatgg agtgggagct caagaaggaa
     2881 aactccaaga ggtggaaccc agagatccag tacacaaaca actacaacga cccccagttt
     2941 gtggactttg ccccggacag caccggggaa tacagaacca ccagacctat cggaacccga
     3001 taccttaccc gaccccttta ataataaccg ggcaggccat gtctgcccgt atttcgcgta
     3061 aggaaatcca ttatgtacta tttaaaaaac acaaactttt ggatgttcgg tttattcttt
     3121 ttcttttact tttttatcat gggagcctac ttcccgtttt tcccgatttg gctacatgac
     3181 atcaaccata tcagcaaaag tgatacgggt attatttttg ccgctatttc tctgttctcg
     3241 ctattattcc aaccgctgtt tggtctgctt tctgacaaac tcggcctcga ctctaggcgg
     3301 ccgcggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3361 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3421 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3481 gggggaggtg tgggaggttt tttcggatcc tctagagtcg acctgcaggc atgcaagctt
     3541 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3601 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3661 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3721 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3781 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3841 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3901 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3961 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4021 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4081 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4141 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4201 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4261 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4381 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4561 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4621 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4681 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4741 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4801 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac ctcgagagcc
     4861 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4921 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4981 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5041 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5101 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5161 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5221 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5281 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5341 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5401 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5461 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5521 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5581 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5641 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5701 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5761 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5821 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5881 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5941 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6001 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6061 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6121 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
     6181 tgcatgcctg caggt
//
