LOCUS       sequence-116341-        9286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 
            rep gene and polycistronic AAV-2 cap gene in Sf9 insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9286)
  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 9286)
  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: pSR657
FEATURES             Location/Qualifiers
     source          1..9286
                     /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(6551..6660)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6684..6775
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6700..6717
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    8876..9010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8913..8932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8967..8986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9039..9204
                     /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 agcttacaga ttacgagtca
     4321 ggtatctggt gccaatgggg cgaggctctg aatacacgcc attagtgtcc acagtaaagt
     4381 ccacattaac agacttgttg tagttggaag tgtactgaat ttcgggattc cagcgtttgc
     4441 tgttttcctt ctgcagctcc cactcgatct ccacgctgac ctgtcccgtg gagtactgtg
     4501 tgatgaagga agcaaacttt gccgcactga aggtggtcga aggattcgca ggtaccgggg
     4561 tgttcttgat gagaatctgt ggaggagggt gtttaagtcc gaatccaccc atgaggggag
     4621 aggggtgaaa atgtccgtcc gtgtgtggaa tctttgccca gatgggcccc tgaaggtaca
     4681 catctctgtc ctgccagacc atgcctggaa gaacgccttg tgtgttgaca tctgcggtag
     4741 ctgcttgtct gttgcctctc tggaggttgg tagatacaga accatactgc tccgtagcca
     4801 cgggattggt tgtcctgatt tcctcttcgt ctgtaatcat gaccttttca atgtccacat
     4861 ttgttttctc tgagccttgc ttcccaaaga tgagaacccc gctctgagga aaaaactttt
     4921 cttcatcgtc cttgtggctt gccatggccg ggcccggatt caccagagag tctctgccat
     4981 tgaggtggta cttggtagct ccagtccacg agtattcact gttgttgtta tccgcagatg
     5041 tctttgatac tcgctgctgg cggtaacagg gtccaggaag ccagttccta gactggtccc
     5101 gaatgtcact cgctccggcc tgagaaaact gaagccttga ctgcgtggtg gttccacttg
     5161 gagtgtttgt tctgctcaag taatacaggt actggtcgat gagaggattc atgagacggt
     5221 ccagactctg gctgtgagcg tagctgctgt ggaaaggaac gtcctcaaaa gtgtagctga
     5281 aggtaaagtt gtttccggta cgcagcatct gagaaggaaa gtactccagg cagtaaaatg
     5341 aagagcgtcc tactgcctga ctcccgttgt tcagggtgag gtatccatac tgtggcacca
     5401 tgaagacgtc tgctgggaac ggcgggaggc atccttgatg cgccgagccg aggacgtacg
     5461 ggagctggta ctccgagtca gtaaacacct gaaccgtgct ggtaaggtta ttggcaatcg
     5521 tcgtcgtacc gtcattctgc gtgacctctt tgacttgaat gttaaagagc ttgaagttga
     5581 gtctcttggg tcggaatccc cagttgttgt tgatgagtct ttgccagtca cgtggtgaaa
     5641 agtggcagtg gaatctgttg aagtcaaaat acccccaagg ggtgctgtag ccaaagtagt
     5701 gattgtcgtt cgaggctcct gattggctgg aaatttgttt gtagaggtgg ttgttgtagg
     5761 tgggcagggc ccaggttcgg gtgctggtgg tgatgactct gtcgcccatc catgtggaat
     5821 cgcaatgcca atttcccgag gaattaccca ctccgtcggc gccctcgtta ttgtctgcca
     5881 ttggtgcgcc actgcctgta gccatcgtat tagttcccag accagagggg gctgctggtg
     5941 gctgtccgag aggctggggg tcaggtactg agtctgcgtc tccagtctga ccaaaattca
     6001 atctttttct tgcaggctgc tggcccgcct ttccggttcc cgaggaggag tctggctcca
     6061 caggagagtg ctctaccggc ctcttttttc ccggagccgt cttaacaggt tcctcaacca
     6121 ggcccagagg ttcaagaacc ctctttttcg cctggaagac tgctcgtccg aggttgcccc
     6181 caaaagacgt atcttcttta aggcgctcct gaaactccgc gtcggcgtgg ttgtacttga
     6241 ggtacgggtt gtctccgctg tcgagctgcc ggtcgtaggc tttgtcgtgc tcgagggccg
     6301 cggcgtctgc ctcgttgacc ggctctccct tgtcgagtcc gttgaagggt ccgaggtact
     6361 tgtacccagg aagcacaaga cccctgctgt cgtccttatg ccgctctgcg ggctttggtg
     6421 gtggtgggcc aggtttgagc ttccaccact gtcttattcc ttcagagaga gtgtcctcga
     6481 gccaatcggg tagataaccg tcggcagccg tcttaacagg atcccccggg tgatcaagtc
     6541 ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata gtattttaat taatatacaa
     6601 atgatttgat aataattctt atttaactat aatatattgt gttgggttga attaaaggtc
     6661 cgtatactcc ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa
     6721 taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc
     6781 ggattattca taccgtccca ccatcgggcg cggatctgcc gccctggcgg ggttttacga
     6841 gattgtgatt aaggtcccca gcgaccttga cgagcatctg cccggcattt ctgacagctt
     6901 tgtgaactgg gtggccgaga aggagtggga gttgccgcca gattctgact tggatctgaa
     6961 tctgattgag caggcacccc tgaccgtggc cgagaagctg cagcgcgact ttctgacgga
     7021 gtggcgccgt gtgagtaagg ccccggaggc ccttttcttt gtgcaatttg agaagggaga
     7081 gagctacttc cacttacacg tgctcgtgga aaccaccggg gtgaaatcct tagttttggg
     7141 acgtttcctg agtcagattc gcgaaaaact gattcagaga atttaccgcg ggatcgagcc
     7201 gactttgcca aactggttcg cggtcacaaa gaccagaaac ggcgccggag gcgggaacaa
     7261 ggtggtggac gagtgctaca tccccaatta cttgctcccc aaaacccagc ctgagctcca
     7321 gtgggcgtgg actaatttag aacagtattt aagcgcctgt ttgaatctca cggagcgtaa
     7381 acggttggtg gcgcagcatc tgacgcacgt gtcgcagacg caggagcaga acaaagagaa
     7441 tcagaatccc aattctgacg cgccggtgat cagatcaaaa acttcagcca ggtacatgga
     7501 gctggtcggg tggctcgtgg acaaggggat tacctcggag aagcagtgga tccaggagga
     7561 ccaggcctca tacatctcct tcaatgcggc ctccaactcg cggtcccaaa tcaaggctgc
     7621 cttggacaat gcgggaaaga ttatgagcct gactaaaacc gcccccgact acctggtggg
     7681 ccagcagccc gtggaggaca tttccagcaa tcggatttat aaaattttgg aactaaacgg
     7741 gtacgatccc caatatgcgg cttccgtctt tctgggatgg gccacgaaaa agttcggcaa
     7801 gaggaacacc atctggctgt ttgggcctgc aactaccggg aagaccaaca tcgcggaggc
     7861 catagcccac actgtgccct tctacgggtg cgtaaactgg accaatgaga actttccctt
     7921 caacgactgt gtcgacaaga tggtgatctg gtgggaggag gggaagatga ccgccaaggt
     7981 cgtggagtcg gccaaagcca ttctcggagg aagcaaggtg cgcgtggacc agaaatgcaa
     8041 gtcctcggcc cagatagacc cgactcccgt gatcgtcacc tccaacacca acatgtgcgc
     8101 cgtgattgac gggaactcaa cgaccttcga acaccagcag ccgttgcaag accggatgtt
     8161 caaatttgaa ctcacccgcc gtctggatca tgactttggg aaggtcacca agcaggaagt
     8221 caaagacttt ttccggtggg caaaggatca cgtggttgag gtggagcatg aattctacgt
     8281 caaaaagggt ggagccaaga aaagacccgc ccccagtgac gcagatataa gtgagcccaa
     8341 acgggtgcgc gagtcagttg cgcagccatc gacgtcagac gcggaagctt cgatcaacta
     8401 cgcagacagg taccaaaaca aatgttctcg tcacgtgggc atgaatctga tgctgtttcc
     8461 ctgcagacaa tgcgagagaa tgaatcagaa ttcaaatatc tgcttcactc acggacagaa
     8521 agactgttta gagtgctttc ccgtgtcaga atctcaaccc gtttctgtcg tcaaaaaggc
     8581 gtatcagaaa ctgtgctaca ttcatcatat catgggaaag gtgccagacg cttgcactgc
     8641 ctgcgatctg gtcaatgtgg atttggatga ctgcatcttt gaacaataaa ctcgaggaat
     8701 caagcttgca tgcctgcagg tcgactctag agcctgcagt ctcgacaagc ttgtcgagaa
     8761 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     8821 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     8881 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     8941 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     9001 tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaatctagt
     9061 tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc
     9121 atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt
     9181 tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa
     9241 acatcctgcc aactccatgt gacaaaccgt catcttcggc tacttt
//
