LOCUS       40924_19756        6978 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Baculovirus expression/transfer vector pFB-6HZB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  AUTHORS   Mahajan P, Savitsky P, Burgess-Brown N, Gileadi O.
  TITLE     Baculovirus expression/transfer vector pFB-6HZB
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6978)
  AUTHORS   Mahajan P, Savitsky P, Burgess-Brown N, Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-DEC-2014) Structural Genomics Consortium, University 
            of Oxford, Old Road Campus Research Building, Roosevelt Drive, 
            Oxford, Oxfordshire OX3 7DQ, UK
REFERENCE   3  (bases 1 to 6978)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6978)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-DEC-2014) 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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             154..171
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             181..351
                     /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             358..378
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQS"
     protein_bind    complement(408..441)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        456..901
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             902..2320
                     /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    2565..2699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  complement(2728..2893)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3077..3532
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3559..3663
                     /label=AmpR promoter
     CDS             3664..4521
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4695..5283
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  complement(5586..5810)
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5880..6410)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(6599..6627)
                     /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 atcatcatca ttcttctggt
      181 gtggataaca agttcaacaa ggagcgtcga agagctcgcc gtgaaattcg ccatctgccg
      241 aacctgaacc gcgaacagcg tcgcgcattt attcgcagcc tgcgcgatga tccgagccag
      301 agcgcgaacc tgctggcgga agcgaagaag ctgaacgatg cgcagccgaa gggtacagag
      361 aacctgtact tccaatccat aagctagctt ctcctcctga aagatccata acttcgtata
      421 gcatacatta tacgaagtta tgcggccgcg acgtccacat atacctgccg ttcactatta
      481 tttagtgaaa tgagatatta tgatattttc tgaattgtga ttaaaaaggc aactttatgc
      541 ccatgcaaca gaaactataa aaaatacaga gaatgaaaag aaacagatag attttttagt
      601 tctttaggcc cgtagtctgc aaatcctttt atgattttct atcaaacaaa agaggaaaat
      661 agaccagttg caatccaaac gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg
      721 gtttgttact gataaagcag gcaagaccta aaatgtgtaa agggcaaagt gtatactttg
      781 gcgtcacccc ttacatattt taggtctttt tttattgtgc gtaactaact tgccatcttc
      841 aaacaggagg gctggaagaa gcagaccgct aacacagtac ataaaaaagg agacatgaac
      901 gatgaacatc aaaaagtttg caaaacaagc aacagtatta acctttacta ccgcactgct
      961 ggcaggaggc gcaactcaag cgtttgcgaa agaaacgaac caaaagccat ataaggaaac
     1021 atacggcatt tcccatatta cacgccatga tatgctgcaa atccctgaac agcaaaaaaa
     1081 tgaaaaatat aaagttcctg agttcgattc gtccacaatt aaaaatatct cttctgcaaa
     1141 aggcctggac gtttgggaca gctggccatt acaaaacact gacggcactg tcgcaaacta
     1201 tcacggctac cacatcgtct ttgcattagc cggagatcct aaaaatgcgg atgacacatc
     1261 gatttacatg ttctatcaaa aagtcggcga aacttctatt gacagctgga aaaacgctgg
     1321 ccgcgtcttt aaagacagcg acaaattcga tgcaaatgat tctatcctaa aagaccaaac
     1381 acaagaatgg tcaggttcag ccacatttac atctgacgga aaaatccgtt tattctacac
     1441 tgatttctcc ggtaaacatt acggcaaaca aacactgaca actgcacaag ttaacgtatc
     1501 agcatcagac agctctttga acatcaacgg tgtagaggat tataaatcaa tctttgacgg
     1561 tgacggaaaa acgtatcaaa atgtacagca gttcatcgat gaaggcaact acagctcagg
     1621 cgacaaccat acgctgagag atcctcacta cgtagaagat aaaggccaca aatacttagt
     1681 atttgaagca aacactggaa ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa
     1741 agcatactat ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag
     1801 cgataaaaaa cgcacggctg agttagcaaa cggcgctctc ggtatgattg agctaaacga
     1861 tgattacaca ctgaaaaaag tgatgaaacc gctgattgca tctaacacag taacagatga
     1921 aattgaacgc gcgaacgtct ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg
     1981 cggatcaaaa atgacgattg acggcattac gtctaacgat atttacatgc ttggttatgt
     2041 ttctaattct ttaactggcc catacaagcc gctgaacaaa actggccttg tgttaaaaat
     2101 ggatcttgat cctaacgatg taacctttac ttactcacac ttcgctgtac ctcaagcgaa
     2161 aggaaacaat gtcgtgatta caagctatat gacaaacaga ggattctacg cagacaaaca
     2221 atcaacgttt gcgcctagct tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa
     2281 agacagcatc cttgaacaag gacaattaac agttaacaaa taaaaacgca aaagaaaatg
     2341 ccgatatcct attggcattg acgtcaggtg gcacttttcg aggagtttac tagtaagtaa
     2401 aggtggatac ggatccgaat tcgagctccg tcgacaagct tgtcgagaag tactagagga
     2461 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2521 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     2701 gatcactgat atcgcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt
     2761 ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca tctattttgt
     2821 cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt
     2881 ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca
     2941 actccatgtg acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt
     3001 tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc
     3061 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3121 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3181 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3241 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3301 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3361 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3421 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3481 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcaggtggc
     3541 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3601 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3661 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3721 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3781 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3841 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3901 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3961 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4021 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4081 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4141 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4201 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4261 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4321 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4381 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4441 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4501 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4561 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4621 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4681 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4741 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4801 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4861 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4921 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4981 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5041 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     5101 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5161 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5221 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5281 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5341 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5401 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5461 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaga
     5521 ccagccgcgt aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa
     5581 gcgggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat
     5641 aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa
     5701 agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc
     5761 ccgtcgtatt aaagaggggc gtggccaagg gcatggtaaa gactatattc gcggcgttgt
     5821 gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag
     5881 gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta
     5941 tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca
     6001 agcgcgttgg cctcatgctt gagcagattg atgagcgcgg tggcaatgcc ctgcctccgg
     6061 tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacctg
     6121 ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg
     6181 atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag
     6241 taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg
     6301 acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt
     6361 gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct
     6421 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     6481 agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
     6541 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
     6601 tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt
     6661 gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa
     6721 cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta
     6781 cggcaaggtg ctgtgcacgg atctgccctg gcttcaggag atcggaagac ctcggccgtc
     6841 gcggcgcttg ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga
     6901 aggcgagcat cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat
     6961 actccggaat attaatag
//
