LOCUS       sequence-351927-        7427 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 7427)
  AUTHORS   Abe K
  TITLE     Vectors for baculovirus gene transfer into Mammalian cells to 
            produce recombinant proteins.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7427)
  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            "Unpublished"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: pFastBacMam5_N_Flag_His8_eGFP_3C
FEATURES             Location/Qualifiers
     source          1..7427
                     /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="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1775..1798
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             join(1799..1801,1802..1804,1805..2515)
                     /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(1844..1865)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2105..2124)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2452..2473
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2525..2548
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    2597..3185
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2650..2670)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3068..3079)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3197..3318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3452..3543)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(3510..3527)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        join(3895..3900,3901..3917,3918..3923)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4093..4626
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4693..4917
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(5051..5068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5222..5810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5302..5321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5981..6772,6773..6841))
                     /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     6604..6623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6842..6946)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6972..7426)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7109..7130)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7320..7339
                     /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 gatcgccggc accatgggtg actacaagga
     1741 cgacgacgac aagagcagcg gcatgggcag cagccatcat catcatcatc accaccacat
     1801 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1861 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1921 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1981 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2041 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2101 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2161 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2221 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2281 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2341 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2401 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2461 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagagcag
     2521 cggcttggaa gttctgtttc aagggccagc tagcgaattc gtcgactcta gactcgaggc
     2581 atgcgcggcc gcatagaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2641 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2701 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2761 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2821 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2881 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2941 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     3001 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3061 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3121 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3181 cccgcctgag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3241 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3301 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3361 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtag gatccgcgcc
     3421 cgatggtggg acggtatgaa taatccggaa tatttatagg tttttttatt acaaaactgt
     3481 tacgaaaaca gtaaaatact tatttatttg cgagatggtt atcattttaa ttatctccat
     3541 gatctattaa tattccggag tatacgtagc caaccactag aactatagct agagtcctgg
     3601 gcgaacaaac gatgctcgcc ttccagaaaa ccgaggatgc gaaccacttc atccggggtc
     3661 agcaccaccg gcaagcgccg cgacggccga ggtcttccga tctcctgaag ccagggcaga
     3721 tccgtgcaca gcaccttgcc gtagaagaac agcaaggccg ccaatgcctg acgatgcgtg
     3781 gagaccgaaa ccttgcgctc gttcgccagc caggacagaa atgcctcgac ttcgctgctg
     3841 cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagttgaca
     3901 taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc
     3961 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     4021 gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     4081 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     4141 acaaagttag gtggctcaag tatgggcatc attcgcacat gtaggctcgg ccctgaccaa
     4201 gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga cgtagccacc
     4261 tactcccaac atcagccgga ctccgattac ctcgggaact tgctccgtag taagacattc
     4321 atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg
     4381 cccaagtttg agcagccgcg tagtgagatc tatatctatg atctcgcagt ctccggcgag
     4441 caccggaggc agggcattgc caccgcgctc atcaatctcc tcaagcatga ggccaacgcg
     4501 cttggtgctt atgtgatcta cgtgcaagca gattacggtg acgatcccgc agtggctctc
     4561 tatacaaagt tgggcatacg ggaagaagtg atgcactttg atatcgaccc aagtaccgcc
     4621 acctaacaat tcgttcaagc cgagatcggc ttcccggccg cggagttgtt cggtaaattg
     4681 tcacaacgcc gcgaatatag tctttaccat gcccttggcc acgcccctct ttaatacgac
     4741 gggcaatttg cacttcagaa aatgaagagt ttgctttagc cataacaaaa gtccagtatg
     4801 ctttttcaca gcataactgg actgatttca gtttacaact attctgtcta gtttaagact
     4861 ttattgtcat agtttagatc tattttgttc agtttaagac tttattgtcc gcccacaccc
     4921 gcttacgcag ggcatccatt tattactcaa ccgtaaccga ttttgccagg ttacgcggct
     4981 ggtctatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     5041 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5101 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5161 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5221 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5281 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5341 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5401 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5461 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5521 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5581 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5641 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5701 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5821 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5881 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5941 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6001 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6061 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6121 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6181 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6241 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6301 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6361 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6421 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6481 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6541 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6601 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6661 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6721 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6781 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6841 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6901 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6961 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     7021 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     7081 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaagaac
     7141 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     7201 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7261 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7321 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7381 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtc
//
