LOCUS       62056_10785        7449 bp DNA     circular SYN 10-NOV-2021
DEFINITION  Synthetic construct DNA, expression vector pFastBacMam4, complete 
            sequence.
ACCESSION   LC658351
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7449)
  AUTHORS   Nakanishi H, Suzuki H, Abe K.
  TITLE     Vectors for baculovirus gene transfer into Mammalian cells to 
            product recombinant Flag-His8-eGFP-TEV fusion proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7449)
  AUTHORS   Nakanishi H, Suzuki H, Abe K.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-NOV-2021) Contact:Hanayo Nakanishi Nagoya University, 
            Cellular and Structural Physiology Institute; Furo-cho, Chikusa-ku, 
            Nagoya, Aichi 4648601, Japan
REFERENCE   3  (bases 1 to 7449)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-NOV-2021) Contact:Hanayo Nakanishi Nagoya University, Cellular 
            and Structural Physiology Institute; Furo-cho, Chikusa-ku, Nagoya, 
            Aichi 4648601, Japan"
FEATURES             Location/Qualifiers
     source          1..7449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  183..348
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    complement(377..511)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     536..557
                     /label=pFastBac primer R1
                     /note="pFastBac primer R1"
     enhancer        797..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     1376..1400
                     /label=LNCX_primer
                     /note="LNCX_primer"
     intron          1515..1647
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1692..1710
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1730..1753
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1775..1798
                     /label=8xHis
                     /note="8xHis affinity tag"
     CDS             1799..2515
                     /label=mEGFP
                     /note="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     CDS             2525..2545
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    2597..2599
                     /label=stop codon
                     /note="stop codon"
     misc_feature    2600..3188
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(3200..3321)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3455..3546)
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     promoter        3898..3926
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4115..4645
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     mobile_element  4715..4939
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      complement(5242..5830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6004..6861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6862..6966)
                     /label=AmpR promoter
     rep_origin      complement(6993..7448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
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 cgcagcagca acgatgttac
     4141 gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac
     4201 atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg
     4261 tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa
     4321 cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg
     4381 cgctctcgcg gcttacgttc tgcccaggtt tgagcagccg cgtagtgaga tctatatcta
     4441 tgatctcgca gtctccggcg agcaccggag gcagggcatt gccaccgcgc tcatcaatct
     4501 cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg
     4561 tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt
     4621 tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttcccggc
     4681 cgcggagttg ttcggtaaat tgtcacaacg ccgcgaatat agtctttacc atgcccttgg
     4741 ccacgcccct ctttaatacg acgggcaatt tgcacttcag aaaatgaaga gtttgcttta
     4801 gccataacaa aagtccagta tgctttttca cagcataact ggactgattt cagtttacaa
     4861 ctattctgtc tagtttaaga ctttattgtc atagtttaga tctattttgt tcagtttaag
     4921 actttattgt ccgcccacac ccgcttacgc agggcatcca tttattactc aaccgtaacc
     4981 gattttgcca ggttacgcgg ctggtctgcg gtgtgaaata ccgcacagat gcgtaaggag
     5041 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5101 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5401 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     5461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5641 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6961 tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata ttttgttaaa
     7021 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     7081 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     7141 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     7201 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     7261 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     7321 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     7381 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     7441 gggcgcgtc
//
