LOCUS       40924_19776        6990 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Baculovirus expression/transfer vector pFB-ZB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  AUTHORS   Savitsky P, Burgess-Brown N, Gileadi O.
  TITLE     Baculovirus expression/transfer vector pFB-ZB
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6990)
  AUTHORS   Savitsky P, Burgess-Brown N, Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-MAR-2011) Structural Genomics Consortium, University 
            of Oxford, Old Road Campus Research Building, Roosevelt Drive, 
            Oxford, Oxfordshire OX3 7DQ, UK
REFERENCE   3  (bases 1 to 6990)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6990)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-MAR-2011) Structural Genomics Consortium, University of Oxford, 
            Old Road Campus Research Building, Roosevelt Drive, Oxford, 
            Oxfordshire OX3 7DQ, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             154..180
                     /codon_start=1
                     /label=9xHis
                     /note="9xHis affinity tag"
                     /translation="HHHHHHHHH"
     CDS             193..363
                     /codon_start=1
                     /label=Z-basic tag
                     /note="Z domain of staphylococcal protein A, engineered to 
                     contain multiple positive charges (Hedhammar and Hober, 
                     2007)"
                     /translation="VDNKFNKERRRARREIRHLPNLNREQRRAFIRSLRDDPSQSANLL
                     AEAKKLNDAQPK"
     CDS             370..390
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQS"
     protein_bind    complement(420..453)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        468..913
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             914..2332
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     polyA_signal    2577..2711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  complement(2740..2905)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3089..3544
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3571..3675
                     /label=AmpR promoter
     CDS             3676..4533
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4707..5295
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  complement(5598..5822)
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5892..6422)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(6611..6639)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atctcggtcc gaaaaccatg ggccaccatc accatcacca tcatcatcat
      181 cattcttctg gtgtggataa caagttcaac aaggagcgtc gaagagctcg ccgtgaaatt
      241 cgccatctgc cgaacctgaa ccgcgaacag cgtcgcgcat ttattcgcag cctgcgcgat
      301 gatccgagcc agagcgcgaa cctgctggcg gaagcgaaga agctgaacga tgcgcagccg
      361 aagggtacag agaacctgta cttccaatcc ataagctagc ttctcctcct gaaagatcca
      421 taacttcgta tagcatacat tatacgaagt tatgcggccg cgacgtccac atatacctgc
      481 cgttcactat tatttagtga aatgagatat tatgatattt tctgaattgt gattaaaaag
      541 gcaactttat gcccatgcaa cagaaactat aaaaaataca gagaatgaaa agaaacagat
      601 agatttttta gttctttagg cccgtagtct gcaaatcctt ttatgatttt ctatcaaaca
      661 aaagaggaaa atagaccagt tgcaatccaa acgagagtct aatagaatga ggtcgaaaag
      721 taaatcgcgc gggtttgtta ctgataaagc aggcaagacc taaaatgtgt aaagggcaaa
      781 gtgtatactt tggcgtcacc ccttacatat tttaggtctt tttttattgt gcgtaactaa
      841 cttgccatct tcaaacagga gggctggaag aagcagaccg ctaacacagt acataaaaaa
      901 ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa gcaacagtat taacctttac
      961 taccgcactg ctggcaggag gcgcaactca agcgtttgcg aaagaaacga accaaaagcc
     1021 atataaggaa acatacggca tttcccatat tacacgccat gatatgctgc aaatccctga
     1081 acagcaaaaa aatgaaaaat ataaagttcc tgagttcgat tcgtccacaa ttaaaaatat
     1141 ctcttctgca aaaggcctgg acgtttggga cagctggcca ttacaaaaca ctgacggcac
     1201 tgtcgcaaac tatcacggct accacatcgt ctttgcatta gccggagatc ctaaaaatgc
     1261 ggatgacaca tcgatttaca tgttctatca aaaagtcggc gaaacttcta ttgacagctg
     1321 gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc gatgcaaatg attctatcct
     1381 aaaagaccaa acacaagaat ggtcaggttc agccacattt acatctgacg gaaaaatccg
     1441 tttattctac actgatttct ccggtaaaca ttacggcaaa caaacactga caactgcaca
     1501 agttaacgta tcagcatcag acagctcttt gaacatcaac ggtgtagagg attataaatc
     1561 aatctttgac ggtgacggaa aaacgtatca aaatgtacag cagttcatcg atgaaggcaa
     1621 ctacagctca ggcgacaacc atacgctgag agatcctcac tacgtagaag ataaaggcca
     1681 caaatactta gtatttgaag caaacactgg aactgaagat ggctaccaag gcgaagaatc
     1741 tttatttaac aaagcatact atggcaaaag cacatcattc ttccgtcaag aaagtcaaaa
     1801 acttctgcaa agcgataaaa aacgcacggc tgagttagca aacggcgctc tcggtatgat
     1861 tgagctaaac gatgattaca cactgaaaaa agtgatgaaa ccgctgattg catctaacac
     1921 agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg aacggcaaat ggtacctgtt
     1981 cactgactcc cgcggatcaa aaatgacgat tgacggcatt acgtctaacg atatttacat
     2041 gcttggttat gtttctaatt ctttaactgg cccatacaag ccgctgaaca aaactggcct
     2101 tgtgttaaaa atggatcttg atcctaacga tgtaaccttt acttactcac acttcgctgt
     2161 acctcaagcg aaaggaaaca atgtcgtgat tacaagctat atgacaaaca gaggattcta
     2221 cgcagacaaa caatcaacgt ttgcgcctag cttcctgctg aacatcaaag gcaagaaaac
     2281 atctgttgtc aaagacagca tccttgaaca aggacaatta acagttaaca aataaaaacg
     2341 caaaagaaaa tgccgatatc ctattggcat tgacgtcagg tggcactttt cgaggagttt
     2401 actagtaagt aaaggtggat acggatccga attcgagctc cgtcgacaag cttgtcgaga
     2461 agtactagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2521 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2581 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2641 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2701 atgtctggat ctgatcactg atatcgccta ggagatccga accagataag tgaaatctag
     2761 ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc
     2821 catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag
     2881 ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca
     2941 aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctactttttc tctgtcacag
     3001 aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg
     3061 cttatttgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     3121 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3181 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3241 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3301 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3361 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3421 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3481 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3541 atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3601 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3661 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3721 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3781 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3841 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3901 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3961 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4021 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4081 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4141 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4201 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4261 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4321 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4381 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4441 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4501 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4561 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4621 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4681 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4741 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4801 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     4861 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4921 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4981 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5041 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
     5101 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5161 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5221 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5281 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5341 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5401 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5461 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     5521 tcacaccgca gaccagccgc gtaacctggc aaaatcggtt acggttgagt aataaatgga
     5581 tgccctgcgt aagcgggtgt gggcggacaa taaagtctta aactgaacaa aatagatcta
     5641 aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt
     5701 atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga
     5761 agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa gggcatggta aagactatat
     5821 tcgcggcgtt gtgacaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga
     5881 acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc ttcccgtatg
     5941 cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg cacgtagatc
     6001 acataagcac caagcgcgtt ggcctcatgc ttgagcagat tgatgagcgc ggtggcaatg
     6061 ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc actacgcggc
     6121 tgctcaaacc tgggcagaac gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag
     6181 gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc ggagtccggc
     6241 tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc aagagcagcc
     6301 cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc catacttgag
     6361 ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac
     6421 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
     6481 tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     6541 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     6601 cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg
     6661 tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc
     6721 ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt
     6781 gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg agatcggaag
     6841 acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct
     6901 cggttttctg gaaggcgagc atcgtttgtt cgcccaggac tctagctata gttctagtgg
     6961 ttggctacgt atactccgga atattaatag
//
