LOCUS       sequence-351924-        7382 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 7382)
  AUTHORS   Abe K
  TITLE     Vectors for baculovirus gene transfer into Mammalian cells to 
            produce recombinant proteins.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7382)
  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            "Unpublished"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: pFastBacMam3_N_His8_eGFP_3C
FEATURES             Location/Qualifiers
     source          1..7382
                     /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..2503
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    2552..3140
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2605..2625)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3023..3034)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3152..3273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3407..3498)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(3465..3482)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        join(3850..3855,3856..3872,3873..3878)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4048..4581
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4648..4872
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(5006..5023)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5177..5765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5257..5276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5936..6727,6728..6796))
                     /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     6559..6578
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6797..6901)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6927..7381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7064..7085)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7275..7294
                     /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 agcagcggct tggaagttct gtttcaaggg ccagctagcg aattcgtcga
     2521 ctctagactc gaggcatgcg cggccgcata gaatcaacct ctggattaca aaatttgtga
     2581 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2641 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2701 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2761 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2821 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2881 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2941 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3001 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3061 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3121 cctttgggcc gcctccccgc ctgagacatg ataagataca ttgatgagtt tggacaaacc
     3181 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3241 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3301 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3361 ggtaggatcc gcgcccgatg gtgggacggt atgaataatc cggaatattt ataggttttt
     3421 ttattacaaa actgttacga aaacagtaaa atacttattt atttgcgaga tggttatcat
     3481 tttaattatc tccatgatct attaatattc cggagtatac gtagccaacc actagaacta
     3541 tagctagagt cctgggcgaa caaacgatgc tcgccttcca gaaaaccgag gatgcgaacc
     3601 acttcatccg gggtcagcac caccggcaag cgccgcgacg gccgaggtct tccgatctcc
     3661 tgaagccagg gcagatccgt gcacagcacc ttgccgtaga agaacagcaa ggccgccaat
     3721 gcctgacgat gcgtggagac cgaaaccttg cgctcgttcg ccagccagga cagaaatgcc
     3781 tcgacttcgc tgctgcccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca
     3841 cgaacccagt tgacataagc ctgttcggtt cgtaaactgt aatgcaagta gcgtatgcgc
     3901 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     3961 ttcatggctt gttatgactg tttttttgta cagtctatgc ctcgggcatc caagcagcaa
     4021 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     4081 ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg
     4141 ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc
     4201 ggagacgtag ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc
     4261 cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc
     4321 gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc
     4381 gcagtctccg gcgagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag
     4441 catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat
     4501 cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc
     4561 gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag
     4621 ttgttcggta aattgtcaca acgccgcgaa tatagtcttt accatgccct tggccacgcc
     4681 cctctttaat acgacgggca atttgcactt cagaaaatga agagtttgct ttagccataa
     4741 caaaagtcca gtatgctttt tcacagcata actggactga tttcagttta caactattct
     4801 gtctagttta agactttatt gtcatagttt agatctattt tgttcagttt aagactttat
     4861 tgtccgccca cacccgctta cgcagggcat ccatttatta ctcaaccgta accgattttg
     4921 ccaggttacg cggctggtct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4981 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5041 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5101 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5161 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5221 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5281 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5341 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5401 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5461 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5521 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5581 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5641 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5701 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5761 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5821 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5881 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5941 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6001 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6061 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6121 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6181 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6241 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6301 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6361 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6421 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6481 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6541 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6601 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6661 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6721 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6781 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6841 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6901 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     6961 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     7021 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     7081 tccactatta agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     7141 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     7201 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     7261 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     7321 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7381 tc
//
