LOCUS       sequence-242820-        4199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes yeast full-length Flp with C-terminal 3xFLAG tag to be 
            expressed in a baculovirus/insect cell expression system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4199)
  AUTHORS   Jensen IS, Inui K, Drakulic S, Jayaprakash S, Sander B, Golas MM
  TITLE     Expression of Flp Protein in a Baculovirus/Insect Cell System for 
            Biotechnological Applications.
  JOURNAL   Protein J. 2017 Aug;36(4):332-342. doi: 10.1007/s10930-017-9724-z.
  PUBMED    28660316
REFERENCE   2  (bases 1 to 4199)
  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"; doi: "10"; journalName: 
            "Protein J"; date: "2017-08"; volume: "36"; issue: "4"; pages: 
            "332-342"
COMMENT     Sequence Label: pGS-BacA-21222-FLP-3xFLAG
FEATURES             Location/Qualifiers
     source          1..4199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  1..166
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      356..944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     845..864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  1079..1302
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1369..1902)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2091..2096,2097..2113,2114..2119))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2190..2223)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2300..2391
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2316..2333
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2472..3740
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFGILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             3747..3812
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3945..4079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3982..4001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4036..4055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct
       61 tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa
      121 ctccatttcc acccctccca gttcccaact attttgtccg cccacaaccg gttgacttgg
      181 gtcaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
      241 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
      301 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      361 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      421 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      481 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      541 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      601 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      661 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      721 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      781 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      841 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      901 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      961 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1021 cccctgattg acttgggtcg ctcttcctgt ggatgcgcag atgccctgcg taagcgggtg
     1081 tgggcggaca ataaagtctt aaactgaaca aaatagatct aaactatgac aataaagtct
     1141 taaactagac agaatagttg taaactgaaa tcagtccagt tatgctgtga aaaagcatac
     1201 tggacttttg ttatggctaa agcaaactct tcattttctg aagtgcaaat tgcccgtcgt
     1261 attaaagagg ggcgtggcca agggcatgta aagactatat tcgcggcgtt gtgacaattt
     1321 accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt
     1381 acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc
     1441 cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt
     1501 ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc
     1561 ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac
     1621 gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt
     1681 cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc
     1741 tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc
     1801 agggccgagc ctacatgtgc gaatgatgcc catacttgag ccacctaact ttgttttagg
     1861 gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac atcgttgctg ctccataaca
     1921 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     1981 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     2041 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa
     2101 ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc
     2161 cggcaacctt gggcagcagc gaagtcgcca taacttcgta tagcatacat tatacgaagt
     2221 tatctgtaac tataacggtc ctaaggtagc gagtttaaac actagtatcg attcgcgacc
     2281 tactccggaa tattaataga tcatggagat aattaaaatg ataaccatct cgcaaataaa
     2341 taagtatttt actgttttcg taacagtttt gtaataaaaa aacctataaa tattccggat
     2401 tattcatacc gtcccaccat cgggcgcgga tcccggtccg aagcgcgcgg aattcaaagg
     2461 cctacgtcga catgccacaa tttggtatat tatgtaaaac accacctaag gtgcttgttc
     2521 gtcagtttgt ggaaaggttt gaaagacctt caggtgagaa aatagcatta tgtgctgctg
     2581 aactaaccta tttatgttgg atgattacac ataacggaac agcaatcaag agagccacat
     2641 tcatgagcta taatactatc ataagcaatt cgctgagttt cgatattgtc aataaatcac
     2701 tccagtttaa atacaagacg caaaaagcaa caattctgga agcctcatta aagaaattga
     2761 ttcctgcttg ggaatttaca attattcctt actatggaca aaaacatcaa tctgatatca
     2821 ctgatattgt aagtagtttg caattacagt tcgaatcatc ggaagaagca gataagggaa
     2881 atagccacag taaaaaaatg cttaaagcac ttctaagtga gggtgaaagc atctgggaga
     2941 tcactgagaa aatactaaat tcgtttgagt atacttcgag atttacaaaa acaaaaactt
     3001 tataccaatt cctcttccta gctactttca tcaattgtgg aagattcagc gatattaaga
     3061 acgttgatcc gaaatcattt aaattagtcc aaaataagta tctgggagta ataatccagt
     3121 gtttagtgac agagacaaag acaagcgtta gtaggcacat atacttcttt agcgcaaggg
     3181 gtaggatcga tccacttgta tatttggatg aatttttgag gaattctgaa ccagtcctaa
     3241 aacgagtaaa taggaccggc aattcttcaa gcaataaaca ggaataccaa ttattaaaag
     3301 ataacttagt cagatcgtac aataaagctt tgaagaaaaa tgcgccttat tcaatctttg
     3361 ctataaaaaa tggcccaaaa tctcacattg gaagacattt gatgacctca tttctttcaa
     3421 tgaagggcct aacggagttg actaatgttg tgggaaattg gagcgataag cgtgcttctg
     3481 ccgtggccag gacaacgtat actcatcaga taacagcaat acctgatcac tacttcgcac
     3541 tagtttctcg gtactatgca tatgatccaa tatcaaagga aatgatagca ttgaaggatg
     3601 agactaatcc aattgaggag tggcagcata tagaacagct aaagggtagt gctgaaggaa
     3661 gcatacgata ccccgcatgg aatgggataa tatcacagga ggtactagac tacctttcat
     3721 cctacataaa tagacgcata tctagagact acaaagacca tgacggtgat tataaagatc
     3781 atgacatcga ttacaaggat gacgatgaca agtaaaagct tgtcgagaag tactagagga
     3841 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3901 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     4081 gatcactgct tgagcctaga agatccggct gctaacaaag cccgaaagga agctgagttg
     4141 gctgctgcca ccgctgagca ataactatca taacccctag ggtataccca tctaattgg
//
