LOCUS       sequence-274696-        3830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flippase-myc fusion ORF with stop codon between attL1-attL2 on 
            Gateway entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3830)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3830)
  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"
COMMENT     Sequence Label: pENTR-Flippase-myc
FEATURES             Location/Qualifiers
     source          1..3830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             674..1942
                     /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="MSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     protein_bind    complement(1955..2054)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2071..2090)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2072..2090)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2095..2111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2095..2111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2213..2232)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2224..3033
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2291..2310)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3180..3768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3669..3688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatgagcc aatttgatat attatgtaaa acaccaccta aggtcctggt
      721 tcgtcagttt gtggaaaggt ttgaaagacc ttcaggggaa aaaatagcat catgtgctgc
      781 tgaactaacc tatttatgtt ggatgattac tcataacgga acagcaatca agagagccac
      841 attcatgagc tataatacta tcataagcaa ttcgctgagt ttcgatattg tcaacaaatc
      901 actccagttt aaatacaaga cgcaaaaagc aacaattctg gaagcctcat taaagaaatt
      961 aattcctgct tgggaattta caattattcc ttacaatgga caaaaacatc aatctgatat
     1021 cactgatatt gtaagtagtt tgcaattaca gttcgaatca tcggaagaag cagataaggg
     1081 aaatagccac agtaaaaaaa tgcttaaagc acttctaagt gagggtgaaa gcatctggga
     1141 gatcactgag aaaatactaa attcgtttga gtatacctcg agatttacaa aaacaaaaac
     1201 tttataccaa ttcctcttcc tagctacttt catcaattgt ggaagattca gcgatattaa
     1261 gaacgttgat ccgaaatcat ttaaattagt ccaaaataag tatctgggag taataatcca
     1321 gtgtttagtg acagagacaa agacaagcgt tagtaggcac atatacttct ttagcgcaag
     1381 gggtaggatc gatccacttg tatatttgga tgaatttttg aggaactctg aaccagtcct
     1441 aaaacgagta aataggaccg gcaattcttc aagcaacaaa caggaatacc aattattaaa
     1501 agataactta gtcagatcgt acaacaaggc tttgaagaaa aatgcgcctt atccaatctt
     1561 tgctataaag aatggcccaa aatctcacat tggaagacat ttgatgacct catttctgtc
     1621 aatgaagggc ctaacggagt tgactaatgt tgtgggaaat tggagcgata agcgtgcttc
     1681 tgccgtggcc aggacaacgt atactcatca gataacagca atacctgatc actacttcgc
     1741 actagtttct cggtactatg catatgatcc aatatcaaag gaaatgatag cattgaagga
     1801 tgagactaat ccaattgagg agtggcagca tatagaacag ctaaagggta gtgctgaagg
     1861 aagcatacga taccccgcat ggaatgggat aatatcacag gaggtactag actacctttc
     1921 atcctacata aatagacgca taggaccggt gtgaacccag ctttcttgta caaagttggc
     1981 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     2041 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     2101 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     2161 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     2221 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     2281 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2341 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     2401 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     2461 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     2521 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     2581 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     2641 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2701 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2761 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2821 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2881 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2941 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3001 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     3061 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     3121 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3181 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3241 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3301 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3361 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3421 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3481 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3541 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3601 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3661 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3721 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3781 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
