LOCUS       sequence-351925-        7430 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 7430)
  AUTHORS   Abe K
  TITLE     Vectors for baculovirus gene transfer into Mammalian cells to 
            produce recombinant proteins.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7430)
  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            "Unpublished"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: pFastBacMam4_N_Flag_His8_eGFP_TEV
FEATURES             Location/Qualifiers
     source          1..7430
                     /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..2545
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    2600..3188
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2653..2673)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3071..3082)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3200..3321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3455..3546)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(3513..3530)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        join(3898..3903,3904..3920,3921..3926)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4096..4629
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4696..4920
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(5054..5071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5225..5813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5305..5324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5984..6775,6776..6844))
                     /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     6607..6626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6845..6949)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6975..7429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7112..7133)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7323..7342
                     /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 cggcgagaac ctgtacttcc agggaggtgg agctagcgaa ttcgtcgact ctagactcga
     2581 ggcatgcgcg gccgcataga atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2641 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2701 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2761 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2821 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2881 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2941 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3001 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3061 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3121 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3181 ctccccgcct gagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3241 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3301 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3361 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taggatccgc
     3421 gcccgatggt gggacggtat gaataatccg gaatatttat aggttttttt attacaaaac
     3481 tgttacgaaa acagtaaaat acttatttat ttgcgagatg gttatcattt taattatctc
     3541 catgatctat taatattccg gagtatacgt agccaaccac tagaactata gctagagtcc
     3601 tgggcgaaca aacgatgctc gccttccaga aaaccgagga tgcgaaccac ttcatccggg
     3661 gtcagcacca ccggcaagcg ccgcgacggc cgaggtcttc cgatctcctg aagccagggc
     3721 agatccgtgc acagcacctt gccgtagaag aacagcaagg ccgccaatgc ctgacgatgc
     3781 gtggagaccg aaaccttgcg ctcgttcgcc agccaggaca gaaatgcctc gacttcgctg
     3841 ctgcccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagttg
     3901 acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg
     3961 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     4021 tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     4081 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
     4141 aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac
     4201 caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc
     4261 acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca
     4321 ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt
     4381 ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc
     4441 gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac
     4501 gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct
     4561 ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc
     4621 gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa
     4681 ttgtcacaac gccgcgaata tagtctttac catgcccttg gccacgcccc tctttaatac
     4741 gacgggcaat ttgcacttca gaaaatgaag agtttgcttt agccataaca aaagtccagt
     4801 atgctttttc acagcataac tggactgatt tcagtttaca actattctgt ctagtttaag
     4861 actttattgt catagtttag atctattttg ttcagtttaa gactttattg tccgcccaca
     4921 cccgcttacg cagggcatcc atttattact caaccgtaac cgattttgcc aggttacgcg
     4981 gctggtctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     5041 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5101 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5161 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5221 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5281 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5341 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5401 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5461 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5521 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5581 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5641 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5701 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5761 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5821 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5881 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5941 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6001 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6061 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6121 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6181 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6241 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6301 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6361 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6421 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6481 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6541 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6601 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6661 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6721 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6781 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6841 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6901 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6961 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     7021 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     7081 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaag
     7141 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     7201 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     7261 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     7321 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     7381 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
//
