LOCUS       sequence-112598-        9284 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 
            rep gene and polycistronic AAV-6 cap gene in Sf9 insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9284)
  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 9284)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pSR646
FEATURES             Location/Qualifiers
     source          1..9284
                     /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(4451..4462)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(6549..6658)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6682..6773
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6698..6715
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    8874..9008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8911..8930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8965..8984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9037..9202
                     /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 agcttacagg ggacgggtga
     4321 ggtaacgggt gccaatgggg cgaggctcag tataaagtcc attgttgtcc acagtgaaat
     4381 caacgttggc agattttgca tagttagatg tatactgcac ttcgggattc cagcgtttgc
     4441 tgttttcttt ctgcagctcc cattcaatct ccacgctcac ttgtcctgtg gaatactggg
     4501 tgatgaatga agcaaacttt gtagccgaaa actctgccgg aggattcgca ggaacaggcg
     4561 tgtttttgat gaggatctga ggaggcgggt gcttaagtcc aaagccgccc atgagaggag
     4621 acgggtgaaa gtgtccatcc gtgtgaggaa ttttggccca aataggaccc tgcaggtata
     4681 cgtctctgtc ttgccacacc attccaggta aggctcccat aacatgcaca tctccggtcg
     4741 cagggtctgt gctgctgctc tggagattga ctgccacagt cccaaatctt tcggtggcca
     4801 cggggttagt ggctttgatt tcctcttcgt ctgtgatcat gacattgtcc aatgcagtgt
     4861 ttgaagctcc ggcgctctcc tttccaaaaa tcatgacacc gctcatggga aagaacttgt
     4921 ctttgtcgtc tttgtgtgag gccatagcag tgccagggtt gattatagat tcacgcccat
     4981 taaggttata ttttgaagca ccagtccagg taaagttgct gttgttgttg tctgtttttg
     5041 ttttagaaac gcgctgctgc cggtaacagg gtccaggtag ccagtttttg ggctgaacag
     5101 acatgccagc tggagacccc cggctaaaca gcaagtcctt gttttgggca cttccggact
     5161 gattctgagt tctgttcagg taatacaggt actggtcgat gagaggattc atcagccggt
     5221 ccaggctctg gctgtgcgcg tagctgctgt ggaaaggcac gtcctcgaag gtgtagctga
     5281 aggtaaagtt attgcccgtt ctcagcatct gcgatgggaa atattccagg cagtaaaagg
     5341 atgaccgtcc cactgcctgg ctgccattgt tgagcgttag gtagccgtac tgcggaatca
     5401 tgaacacgtc cgccgggaac ggagggaggc agccctggtg cgcagagccg aggacgtacg
     5461 gcaactggta ctccgagtcc gagaagactt gaaccgtgct ggtaaggtta ttagcgatgg
     5521 tcgtgacgcc atcattcgtc gtgacctcct tgacttggat gttgaagagc ttgaagttga
     5581 gtctcttggg ccggaatccc caattgttgt tgatgagtcg ctgccagtca cgtggtgaga
     5641 aatggcagtg gaatctgttg aaatcaaaat acccccaggg ggtgctgtag ccgaagtagt
     5701 ggttgtcgtt gctggccccc gttgaagcac tggagatttg cttgtagagg tggttgttat
     5761 aggtgggcaa ggcccatgtt cgggtgctgg tggtgatgac tctgtcgccc agccatgtgg
     5821 aatcgcaatg ccaatttcct gaggcattac ccactccgtc ggcgccttcg ttattgtctg
     5881 ccattggtgc gccaccgcct gaagccattg tagtaggtcc cacagcagcg ggggttgctg
     5941 gaggttctcc gagaggttgt gggtcgggga ctgactctga gtcgccagtc tgaccaaaat
     6001 tgagtctctt tttagcgggc tgctggcctg tcttgccaat gcccgaggag gagtctggct
     6061 cttgtggcga ctgctctacc ggacgtttct ttccaggagc cgtcttagca ccttcctcaa
     6121 ccagaccaaa aggttcgaga accctcttct tggcctggaa gactgctcgc ccgaggttgc
     6181 ccccaaaaga cgtatcttct tgcagacgct cctgaaactc ggcgtcggcg tggttatacc
     6241 gcaggtacgg attgtcaccc gctttgagct gctggtcgta ggccttgtcg tgctcgaggg
     6301 ccgctgcatc cgccgcgttg acgggctccc ccttgtcgag tccgttgaag ggtccgaggt
     6361 acttgtagcc aggaagcacc agaccccggc cgtcgtcctg cttttgctgg ttggctttgg
     6421 gtttcggggc tccaggtttg aggtcccacc actcgcgaat gccctcagag aggttgtcct
     6481 cgagccaatc gggtagataa ccgtcggcag ccaaggcggc ggcccgggtg atcaagtctt
     6541 cgtcgagtga ttgtaaataa aatgtaattt acagtatagt attttaatta atatacaaat
     6601 gatttgataa taattcttat ttaactataa tatattgtgt tgggttgaat taaaggtccg
     6661 tatactccgg aatattaata gatcatggag ataattaaaa tgataaccat ctcgcaaata
     6721 aataagtatt ttactgtttt cgtaacagtt ttgtaataaa aaaacctata aatattccgg
     6781 attattcata ccgtcccacc atcgggcgcg gatctgccgc cctggcgggg ttttacgaga
     6841 ttgtgattaa ggtccccagc gaccttgacg agcatctgcc cggcatttct gacagctttg
     6901 tgaactgggt ggccgagaag gagtgggagt tgccgccaga ttctgacttg gatctgaatc
     6961 tgattgagca ggcacccctg accgtggccg agaagctgca gcgcgacttt ctgacggagt
     7021 ggcgccgtgt gagtaaggcc ccggaggccc ttttctttgt gcaatttgag aagggagaga
     7081 gctacttcca cttacacgtg ctcgtggaaa ccaccggggt gaaatcctta gttttgggac
     7141 gtttcctgag tcagattcgc gaaaaactga ttcagagaat ttaccgcggg atcgagccga
     7201 ctttgccaaa ctggttcgcg gtcacaaaga ccagaaacgg cgccggaggc gggaacaagg
     7261 tggtggacga gtgctacatc cccaattact tgctccccaa aacccagcct gagctccagt
     7321 gggcgtggac taatttagaa cagtatttaa gcgcctgttt gaatctcacg gagcgtaaac
     7381 ggttggtggc gcagcatctg acgcacgtgt cgcagacgca ggagcagaac aaagagaatc
     7441 agaatcccaa ttctgacgcg ccggtgatca gatcaaaaac ttcagccagg tacatggagc
     7501 tggtcgggtg gctcgtggac aaggggatta cctcggagaa gcagtggatc caggaggacc
     7561 aggcctcata catctccttc aatgcggcct ccaactcgcg gtcccaaatc aaggctgcct
     7621 tggacaatgc gggaaagatt atgagcctga ctaaaaccgc ccccgactac ctggtgggcc
     7681 agcagcccgt ggaggacatt tccagcaatc ggatttataa aattttggaa ctaaacgggt
     7741 acgatcccca atatgcggct tccgtctttc tgggatgggc cacgaaaaag ttcggcaaga
     7801 ggaacaccat ctggctgttt gggcctgcaa ctaccgggaa gaccaacatc gcggaggcca
     7861 tagcccacac tgtgcccttc tacgggtgcg taaactggac caatgagaac tttcccttca
     7921 acgactgtgt cgacaagatg gtgatctggt gggaggaggg gaagatgacc gccaaggtcg
     7981 tggagtcggc caaagccatt ctcggaggaa gcaaggtgcg cgtggaccag aaatgcaagt
     8041 cctcggccca gatagacccg actcccgtga tcgtcacctc caacaccaac atgtgcgccg
     8101 tgattgacgg gaactcaacg accttcgaac accagcagcc gttgcaagac cggatgttca
     8161 aatttgaact cacccgccgt ctggatcatg actttgggaa ggtcaccaag caggaagtca
     8221 aagacttttt ccggtgggca aaggatcacg tggttgaggt ggagcatgaa ttctacgtca
     8281 aaaagggtgg agccaagaaa agacccgccc ccagtgacgc agatataagt gagcccaaac
     8341 gggtgcgcga gtcagttgcg cagccatcga cgtcagacgc ggaagcttcg atcaactacg
     8401 cagacaggta ccaaaacaaa tgttctcgtc acgtgggcat gaatctgatg ctgtttccct
     8461 gcagacaatg cgagagaatg aatcagaatt caaatatctg cttcactcac ggacagaaag
     8521 actgtttaga gtgctttccc gtgtcagaat ctcaacccgt ttctgtcgtc aaaaaggcgt
     8581 atcagaaact gtgctacatt catcatatca tgggaaaggt gccagacgct tgcactgcct
     8641 gcgatctggt caatgtggat ttggatgact gcatctttga acaataaact cgaggaatca
     8701 agcttgcatg cctgcaggtc gactctagag cctgcagtct cgacaagctt gtcgagaagt
     8761 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     8821 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     8881 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8941 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     9001 tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga aatctagttc
     9061 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
     9121 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
     9181 ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac
     9241 atcctgccaa ctccatgtga caaaccgtca tcttcggcta cttt
//
