LOCUS       sequence-112889-        9327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 
            rep gene and polycistronic AAV-1 cap gene in Sf9 insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9327)
  AUTHORS   Smith RH, Levy JR, Kotin RM
  TITLE     A simplified baculovirus-AAV expression vector system coupled with 
            one-step affinity purification yields high-titer rAAV stocks from 
            insect cells.
  JOURNAL   Mol Ther. 2009 Nov;17(11):1888-96. doi: 10.1038/mt.2009.128. Epub 
            2009 Jun 16.
  PUBMED    19532142
REFERENCE   2  (bases 1 to 9327)
  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"; doi: 
            "10.1038/mt.2009.128"; journalName: "Mol Ther"; date: "2009-11"; 
            volume: "17"; issue: "11"; pages: "1888-96"
COMMENT     Sequence Label: pSR651
FEATURES             Location/Qualifiers
     source          1..9327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     399..420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        584..688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(689..757,758..1549)
                     /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(907..926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1720..2308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2209..2228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2462..2479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4161..4209
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4234..4253
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4500..4511)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(6592..6701)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6725..6816
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6741..6758
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    8917..9051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8954..8973)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9008..9027
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9080..9245
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      541 attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2101 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2461 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2521 tctgtgcggt atttcacacc gcagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcct acgtacccgt agtggctatg gcagggcttg ccgccccgac
     4021 gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga
     4081 aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg
     4141 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt
     4201 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     4261 tcccccatct cccggtaccg catgctatgc atcagctgct agattcgaaa gcggccgcca
     4321 gtgtgatgga tatctgcaca attcgccctt gcttacaggg gacgggtaag gtaacgggtg
     4381 ccaatggggc gaggctcagt ataaagtcca ttgttgtcca cagtaaaatc aacgttggca
     4441 gattttgcat aattggatgt gtactgcact tcgggattcc agcgcttgct gttttctttc
     4501 tgcagctccc attcaatttc cacactcact tgtcctgtgg agtattgggt gatgaatgaa
     4561 gcaaactttg tagctgaaaa ctccgccgga ggattcgcag gaacaggcgt gtttttgatg
     4621 aggatctgag gaggcgggtt cttgagtcca aagccgccca taagaggaga cgggtgaaag
     4681 tgtccatctg tgtgaggaat tttggcccaa atgggaccct gcaggtacac gtctctatct
     4741 tgccacacca tgccaggtaa tgctcccata gcatgcacat ctccggtcgc agggtctgtg
     4801 ctgctgctct ggaaattgac tgccacggtc ccaaatcttt cggtggccac agggttagtg
     4861 gctttaattt cctcttcgtc tgtaatcatg acattgtcca atgcagtgtt tgaagctccg
     4921 gcgctctctt ttccaaaaat catgacaccg ctcatgggaa agaacttgtc ttcgtcgtct
     4981 ttgtgtgagg ccatagcagt gccagggttg atgatggatt cacgcccatt gaggttatat
     5041 tttgaagcac cagtccaggt aaaattgctg ttgttgttgt ctgtttttgt tttagaaacg
     5101 cgctgctgcc gataacaggg tccaggtagc cagtttttgg gctgaacaga catgccagct
     5161 ggagacccac ggctaaacag caagtccttg ttttgggcac ttccggactg attttgagtt
     5221 ctgttcaggt aatacaggta ttggtcgatg agaggattca tcagccggtc caggctctgg
     5281 ctgtgcgcgt agctgctgtg gaaaggcact tcctcaaagg tgtagctgaa ggtaaagttg
     5341 ttgcccgttc tcagcatctg agaagggaaa tattccaggc agtaaaagga tgaacgtccc
     5401 acggcttggc tgccattgtt gagcgtcagg tagccgtatt gcggaatcat gaacacgtcc
     5461 gccgggaacg gagggaggca gccctggtgc gcagagccga ggacgtacgg aagctggtac
     5521 tccgagtccg agaagacttg aaccgtgctg gtaaggttat tagcgatggt tgtgacgcca
     5581 tcattcgtcg tgacctcctt gacttggatg ttgaagagtt tgaagttgag tctcttgggc
     5641 cggaatcccc aattgttgtt gatgagtcgc tgccagtcac gtggtgaaaa gtggcagtgg
     5701 aatctgttga aatcaaaata cccccagggg gtgctgtagc cgaagtagtg gttgtcgttg
     5761 ctggcccccg ttgaagcact ggagatttgc ttgtagaggt ggttattgta ggtgggcaag
     5821 gcccaggtgc gggtgctggt ggtgatgact ctgtcgccca gccatgtgga atcgcaatgc
     5881 caatttcctg aggcattacc cactccgtcg gcgccttcgt tattgtctgc cattggtgcg
     5941 ccaccgcctg aagccattgt agtaggtccc acagcagcgg gggttgctgg aggttctccg
     6001 agaggttgtg gatcggggac tgactctgag tcgccagtct gaccaaaatt gagtctcttt
     6061 ttagcgggct gctggcctgt cttgccgatg cccgaggagg agtctggctc ttgtggcgac
     6121 tgctctaccg gacgtttctt tccaggagcc gtcttagcgc cttcctcaac cagaccgaga
     6181 ggttcgagaa cccgcttctt ggcctggaag actgctcgcc cgaggttgcc cccaaaagac
     6241 gtatcttctt gcagacgctc ctgaaactcg gcgtcggcgt ggttataccg caggtacgga
     6301 ttgtcacccg ctttgagctg ctggtcgtag gccttgtcgt gctcgagggc cgctgcgtcc
     6361 gccgcgttga cgggctcccc cttgtcgagt ccgttgaagg gtccgaggta cttgtagcca
     6421 ggaagcacca gaccccggcc gtcgtcctgc ttttgctggt tggctttggg cttcggggct
     6481 ccaggtttca agtcccacca ctcgcgaatg ccctcagaga ggttgtcctc gagccaatcg
     6541 ggtagataac cgtcggcagc cgtctttaac aggatcaagt cttcgtcgag tgattgtaaa
     6601 taaaatgtaa tttacagtat agtattttaa ttaatataca aatgatttga taataattct
     6661 tatttaacta taatatattg tgttgggttg aattaaaggt ccgtatactc cggaatatta
     6721 atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt attttactgt
     6781 tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc
     6841 accatcgggc gcggatctgc cgccctggcg gggttttacg agattgtgat taaggtcccc
     6901 agcgaccttg acgagcatct gcccggcatt tctgacagct ttgtgaactg ggtggccgag
     6961 aaggagtggg agttgccgcc agattctgac ttggatctga atctgattga gcaggcaccc
     7021 ctgaccgtgg ccgagaagct gcagcgcgac tttctgacgg agtggcgccg tgtgagtaag
     7081 gccccggagg cccttttctt tgtgcaattt gagaagggag agagctactt ccacttacac
     7141 gtgctcgtgg aaaccaccgg ggtgaaatcc ttagttttgg gacgtttcct gagtcagatt
     7201 cgcgaaaaac tgattcagag aatttaccgc gggatcgagc cgactttgcc aaactggttc
     7261 gcggtcacaa agaccagaaa cggcgccgga ggcgggaaca aggtggtgga cgagtgctac
     7321 atccccaatt acttgctccc caaaacccag cctgagctcc agtgggcgtg gactaattta
     7381 gaacagtatt taagcgcctg tttgaatctc acggagcgta aacggttggt ggcgcagcat
     7441 ctgacgcacg tgtcgcagac gcaggagcag aacaaagaga atcagaatcc caattctgac
     7501 gcgccggtga tcagatcaaa aacttcagcc aggtacatgg agctggtcgg gtggctcgtg
     7561 gacaagggga ttacctcgga gaagcagtgg atccaggagg accaggcctc atacatctcc
     7621 ttcaatgcgg cctccaactc gcggtcccaa atcaaggctg ccttggacaa tgcgggaaag
     7681 attatgagcc tgactaaaac cgcccccgac tacctggtgg gccagcagcc cgtggaggac
     7741 atttccagca atcggattta taaaattttg gaactaaacg ggtacgatcc ccaatatgcg
     7801 gcttccgtct ttctgggatg ggccacgaaa aagttcggca agaggaacac catctggctg
     7861 tttgggcctg caactaccgg gaagaccaac atcgcggagg ccatagccca cactgtgccc
     7921 ttctacgggt gcgtaaactg gaccaatgag aactttccct tcaacgactg tgtcgacaag
     7981 atggtgatct ggtgggagga ggggaagatg accgccaagg tcgtggagtc ggccaaagcc
     8041 attctcggag gaagcaaggt gcgcgtggac cagaaatgca agtcctcggc ccagatagac
     8101 ccgactcccg tgatcgtcac ctccaacacc aacatgtgcg ccgtgattga cgggaactca
     8161 acgaccttcg aacaccagca gccgttgcaa gaccggatgt tcaaatttga actcacccgc
     8221 cgtctggatc atgactttgg gaaggtcacc aagcaggaag tcaaagactt tttccggtgg
     8281 gcaaaggatc acgtggttga ggtggagcat gaattctacg tcaaaaaggg tggagccaag
     8341 aaaagacccg cccccagtga cgcagatata agtgagccca aacgggtgcg cgagtcagtt
     8401 gcgcagccat cgacgtcaga cgcggaagct tcgatcaact acgcagacag gtaccaaaac
     8461 aaatgttctc gtcacgtggg catgaatctg atgctgtttc cctgcagaca atgcgagaga
     8521 atgaatcaga attcaaatat ctgcttcact cacggacaga aagactgttt agagtgcttt
     8581 cccgtgtcag aatctcaacc cgtttctgtc gtcaaaaagg cgtatcagaa actgtgctac
     8641 attcatcata tcatgggaaa ggtgccagac gcttgcactg cctgcgatct ggtcaatgtg
     8701 gatttggatg actgcatctt tgaacaataa actcgaggaa tcaagcttgc atgcctgcag
     8761 gtcgactcta gagcctgcag tctcgacaag cttgtcgaga agtactagag gatcataatc
     8821 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     8881 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     8941 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     9001 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg
     9061 cttgagccta ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt
     9121 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     9181 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     9241 ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg
     9301 tgacaaaccg tcatcttcgg ctacttt
//
