LOCUS       sequence-115626-        9362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 
            rep gene and polycistronic AAV-8 cap gene in Sf9 insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9362)
  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 9362)
  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: pSR660
FEATURES             Location/Qualifiers
     source          1..9362
                     /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(4507..4518)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(6627..6736)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6760..6851
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6776..6793
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    8952..9086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8989..9008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9043..9062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9115..9280
                     /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 agccgcagag accaaagttc
     4321 aactgaaacg aatcaaccgg tttattgatt aacaggcaat tacagattac gggtgaggta
     4381 acgggtgcca atggggcggg gttcagagta cacgccttct gtattaacag caaagtccac
     4441 acttgtagat ttgtagtagt tggaggtgta ctggatctcg gggttccagc gcttgctgtt
     4501 ttccttctgc agctcccatt caatttccac gctgacctgt ccggtgctgt attgcgtgat
     4561 gaaagagttc agctttgact ggttgaaggt ggtcggagga tccgcaggta caggcgtgtt
     4621 cttgatcagg atctgaggcg gaggatgttt caggccaaag ccgcccatca gcggagacgg
     4681 gtggaagttg ccgtccgtgt gaggaatctt ggcccagatg ggaccctgca ggtacacgtc
     4741 ccggttctgc cagaccatac cgggtaaggc cccctggctg ttgacagttc caatttgagg
     4801 agccgtgttt tgctgctgca agttatctgc cacgataccg tattcctctg tagccacagg
     4861 gttagtggtt ttgatttctt cctcgctggt gagcatgaca tcgctgtaat ccgcattgtc
     4921 tctggcagca ttttgtttgc caaaaatcag gatcccgtta ctgggaaaaa aacgctcctc
     4981 gtcgtctttg tgtgttgcca tagcgatgcc aggattagcc aatgaatttc ttccattcag
     5041 atggtatttg gtcccagcag tccaggcaaa gttgctattg ttgttttgcc cggttgtcgt
     5101 tgagacgcgt tgttggcggt aacagggtcc tggcagccag ttctttgcct gattggccat
     5161 tgtattaggc ccaccttggc tgaagcccag agtctgcgta tttgccgtgc ctcctgttgt
     5221 ttgagtccga gacaagtagt acaggtactg gtcaatcaga ggattcatca gccggtccaa
     5281 gctctggctg tgggcgtagc tgctgtggaa aggcacgtcc tcgaaggtgt aagtaaactg
     5341 gaagttgttg ccggttctca gcatctgcga aggaaagtat tccaggcagt agaaggagga
     5401 gcgtcccacg gcctgactac cgttgttgag tgttaggtag ccgtactggg gaatcatgaa
     5461 cacgtccgcc gggaacggag gcaggcagcc ctggtgggca gagccgagaa cgtacggcag
     5521 ctggtactcc gagtccgtaa acacctggat ggtgctggtg aggttattgg cgatggtctt
     5581 ggtgccttca ttctgcgtga cctccttgac ctggatgttg aagagcttga agctgagtct
     5641 cttgggccgg aatccccagt tgttgttgat gagtcgctgc cagtcacgtg gtgaaaagtg
     5701 gcagtggaat ctgttaaagt caaaataccc ccagggggtg ctgtagccga agtaggtgtt
     5761 gtcgttggtg gctcctcccg atgtcccgtt ggagatttgc ttgtagaggt ggttgttgta
     5821 ggtgggcagg gcccaggttc gggtgctggt ggtgatgact ctgtcgccca gccatgtgga
     5881 atcgcaatgc caatttcccg aggaactacc cactccgtcg gcgccttcgt tattgtctgc
     5941 cattggtgcg ccaccgcctg cagccattgt attaggtccc acaccagagg gcgctgctgg
     6001 aggttctccg agaggttgag ggtctggaac tgactctgag tcgccagtct gaccaaaatt
     6061 gagtcttttt ctggcgggct gttggccttt cttgccgatg cccgtagagg agtctggaga
     6121 acgctggggt gatggctcta ccggtctctt ctttccagga gccgtcttag cgccttcctc
     6181 aaccagaccg agaggttcga gaacccgctt cttggcctgg aagactgctc gcccgaggtt
     6241 gcccccaaaa gacgtatctt cttgcagacg ctcctggaat tcagcgtcgg cgtggttata
     6301 acgaaggtaa gggttgtcgc cggcctgcag ttgctggtcg taagccttat cgtgctcaag
     6361 ggcagcggcg tccgcagcgt tcacgggttc gcccttgtcc agcccgttga agggtcctag
     6421 gtatttgtaa ccggggagaa ccaggccgcg tccgtcgtct tgtttctgtt ggttagcctt
     6481 tggcttagga gcgccgggtt tgagggccca ccactcgcga atgccctcag agaggttgtc
     6541 ctcgagccaa tcgggtagat aaccgtcggc agccgtggcg gcaggaatcc cgcgcgcttc
     6601 cccgggtgat caagtcttcg tcgagtgatt gtaaataaaa tgtaatttac agtatagtat
     6661 tttaattaat atacaaatga tttgataata attcttattt aactataata tattgtgttg
     6721 ggttgaatta aaggtccgta tactccggaa tattaataga tcatggagat aattaaaatg
     6781 ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa
     6841 aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga tctgccgccc
     6901 tggcggggtt ttacgagatt gtgattaagg tccccagcga ccttgacgag catctgcccg
     6961 gcatttctga cagctttgtg aactgggtgg ccgagaagga gtgggagttg ccgccagatt
     7021 ctgacttgga tctgaatctg attgagcagg cacccctgac cgtggccgag aagctgcagc
     7081 gcgactttct gacggagtgg cgccgtgtga gtaaggcccc ggaggccctt ttctttgtgc
     7141 aatttgagaa gggagagagc tacttccact tacacgtgct cgtggaaacc accggggtga
     7201 aatccttagt tttgggacgt ttcctgagtc agattcgcga aaaactgatt cagagaattt
     7261 accgcgggat cgagccgact ttgccaaact ggttcgcggt cacaaagacc agaaacggcg
     7321 ccggaggcgg gaacaaggtg gtggacgagt gctacatccc caattacttg ctccccaaaa
     7381 cccagcctga gctccagtgg gcgtggacta atttagaaca gtatttaagc gcctgtttga
     7441 atctcacgga gcgtaaacgg ttggtggcgc agcatctgac gcacgtgtcg cagacgcagg
     7501 agcagaacaa agagaatcag aatcccaatt ctgacgcgcc ggtgatcaga tcaaaaactt
     7561 cagccaggta catggagctg gtcgggtggc tcgtggacaa ggggattacc tcggagaagc
     7621 agtggatcca ggaggaccag gcctcataca tctccttcaa tgcggcctcc aactcgcggt
     7681 cccaaatcaa ggctgccttg gacaatgcgg gaaagattat gagcctgact aaaaccgccc
     7741 ccgactacct ggtgggccag cagcccgtgg aggacatttc cagcaatcgg atttataaaa
     7801 ttttggaact aaacgggtac gatccccaat atgcggcttc cgtctttctg ggatgggcca
     7861 cgaaaaagtt cggcaagagg aacaccatct ggctgtttgg gcctgcaact accgggaaga
     7921 ccaacatcgc ggaggccata gcccacactg tgcccttcta cgggtgcgta aactggacca
     7981 atgagaactt tcccttcaac gactgtgtcg acaagatggt gatctggtgg gaggagggga
     8041 agatgaccgc caaggtcgtg gagtcggcca aagccattct cggaggaagc aaggtgcgcg
     8101 tggaccagaa atgcaagtcc tcggcccaga tagacccgac tcccgtgatc gtcacctcca
     8161 acaccaacat gtgcgccgtg attgacggga actcaacgac cttcgaacac cagcagccgt
     8221 tgcaagaccg gatgttcaaa tttgaactca cccgccgtct ggatcatgac tttgggaagg
     8281 tcaccaagca ggaagtcaaa gactttttcc ggtgggcaaa ggatcacgtg gttgaggtgg
     8341 agcatgaatt ctacgtcaaa aagggtggag ccaagaaaag acccgccccc agtgacgcag
     8401 atataagtga gcccaaacgg gtgcgcgagt cagttgcgca gccatcgacg tcagacgcgg
     8461 aagcttcgat caactacgca gacaggtacc aaaacaaatg ttctcgtcac gtgggcatga
     8521 atctgatgct gtttccctgc agacaatgcg agagaatgaa tcagaattca aatatctgct
     8581 tcactcacgg acagaaagac tgtttagagt gctttcccgt gtcagaatct caacccgttt
     8641 ctgtcgtcaa aaaggcgtat cagaaactgt gctacattca tcatatcatg ggaaaggtgc
     8701 cagacgcttg cactgcctgc gatctggtca atgtggattt ggatgactgc atctttgaac
     8761 aataaactcg aggaatcaag cttgcatgcc tgcaggtcga ctctagagcc tgcagtctcg
     8821 acaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     8881 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     8941 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     9001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     9061 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
     9121 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
     9181 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
     9241 ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg
     9301 ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact
     9361 tt
//
