LOCUS       sequence-351928-        7385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus gene transfer into Mammalian cells to product 
            recombinant proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7385)
  AUTHORS   Abe K
  TITLE     Vectors for baculovirus gene transfer into Mammalian cells to 
            produce recombinant proteins.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7385)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFastBacMam2_N_His8_eGFP_TEV
FEATURES             Location/Qualifiers
     source          1..7385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  183..348
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    377..511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        796..1175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1176..1379
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1330..1350
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1376..1400
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1515..1647
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1630..1649
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1692..1711
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1692..1710
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1730..1753
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             join(1754..1756,1757..1759,1760..2470)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1799..1820)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2060..2079)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2407..2428
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2480..2500
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    2555..3143
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2608..2628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3026..3037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3155..3276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3410..3501)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(3468..3485)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        join(3853..3858,3859..3875,3876..3881)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4051..4584
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4651..4875
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(5009..5026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5180..5768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5260..5279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5939..6730,6731..6799))
                     /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     6562..6581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6800..6904)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6930..7384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7067..7088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7278..7297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccattcgcca ttcaggctgc aaataagcgt tgatattcag tcaattacaa acattaataa
       61 cgaagagatg acagaaaaat tttcattctg tgacagagaa aaagtagccg aagatgacgg
      121 tttgtcacat ggagttggca ggatgtttga ttaaaaacat aacaggaaga aaaatgcccc
      181 gctgtgggcg gacaaaatag ttgggaactg ggaggggtgg aaatggagtt tttaaggatt
      241 atttagggaa gagtgacaaa atagatggga actgggtgta gcgtcgtaag ctaatacgaa
      301 aattaaaaat gacaaaatag tttggaacta gatttcactt atctggttcg gatctcctag
      361 gctcaagcag tgatcagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
      421 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      481 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      541 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
      601 tatggctgat tatgatcctc tagtacttct cgacaagctt ggtaccccta ggagatcttc
      661 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt
      721 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
      781 tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca attacggggt
      841 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      901 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      961 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1021 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     1081 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     1141 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
     1201 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1261 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     1321 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     1381 gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac agttaaattg
     1441 ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcaga agttggtcgt
     1501 gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     1561 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     1621 acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac agctcttaag
     1681 gctagagtac ttaatacgac tcactatagg gatcgccatg ggcagcagcc atcatcatca
     1741 tcatcaccac cacatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1801 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1861 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1921 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1981 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2041 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2101 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2161 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2221 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2281 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2341 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
     2401 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2461 gctgtacaag agcagcggcg agaacctgta cttccaggga ggtggagcta gcgaattcgt
     2521 cgactctaga ctcgaggcat gcgcggccgc atagaatcaa cctctggatt acaaaatttg
     2581 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2641 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2701 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2761 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2821 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2881 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2941 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3001 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3061 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3121 ctccctttgg gccgcctccc cgcctgagac atgataagat acattgatga gtttggacaa
     3181 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     3241 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3301 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     3361 tgtggtagga tccgcgcccg atggtgggac ggtatgaata atccggaata tttataggtt
     3421 tttttattac aaaactgtta cgaaaacagt aaaatactta tttatttgcg agatggttat
     3481 cattttaatt atctccatga tctattaata ttccggagta tacgtagcca accactagaa
     3541 ctatagctag agtcctgggc gaacaaacga tgctcgcctt ccagaaaacc gaggatgcga
     3601 accacttcat ccggggtcag caccaccggc aagcgccgcg acggccgagg tcttccgatc
     3661 tcctgaagcc agggcagatc cgtgcacagc accttgccgt agaagaacag caaggccgcc
     3721 aatgcctgac gatgcgtgga gaccgaaacc ttgcgctcgt tcgccagcca ggacagaaat
     3781 gcctcgactt cgctgctgcc caaggttgcc gggtgacgca caccgtggaa acggatgaag
     3841 gcacgaaccc agttgacata agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg
     3901 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
     3961 gttttcatgg cttgttatga ctgttttttt gtacagtcta tgcctcgggc atccaagcag
     4021 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     4081 cagggcagtc gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt
     4141 aggctcggcc ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag
     4201 ttcggagacg tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg
     4261 ctccgtagta agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct
     4321 ctcgcggctt acgttctgcc caagtttgag cagccgcgta gtgagatcta tatctatgat
     4381 ctcgcagtct ccggcgagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc
     4441 aagcatgagg ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac
     4501 gatcccgcag tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat
     4561 atcgacccaa gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgcg
     4621 gagttgttcg gtaaattgtc acaacgccgc gaatatagtc tttaccatgc ccttggccac
     4681 gcccctcttt aatacgacgg gcaatttgca cttcagaaaa tgaagagttt gctttagcca
     4741 taacaaaagt ccagtatgct ttttcacagc ataactggac tgatttcagt ttacaactat
     4801 tctgtctagt ttaagacttt attgtcatag tttagatcta ttttgttcag tttaagactt
     4861 tattgtccgc ccacacccgc ttacgcaggg catccattta ttactcaacc gtaaccgatt
     4921 ttgccaggtt acgcggctgg tctatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4981 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6901 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     6961 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7021 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7081 gagtccacta ttaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     7141 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     7201 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     7261 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     7321 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     7381 gcgtc
//
