LOCUS       sequence-220037-        5062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse codon-optimized FLP recombinase with bovine 
            growth hormone polyadenylation (bpA) sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5062)
  AUTHORS   Raymond CS, Soriano P
  TITLE     High-efficiency FLP and PhiC31 site-specific recombination in 
            mammalian cells.
  JOURNAL   PLoS One. 2007 Jan 17;2(1):e162. doi: 10.1371/journal.pone.0000162.
  PUBMED    17225864
REFERENCE   2  (bases 1 to 5062)
  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"; journalName: "PLoS One.";
            date: "2007-01-17"; volume: "2(1):e162. doi"; pages: " 
            10.1371/journal.pone.0000162"
COMMENT     Sequence Label: pPGKFLPobpA
FEATURES             Location/Qualifiers
     source          1..5062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(264..269,270..287,288..294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        363..381
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     409..425
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     410..426
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        451..950
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(486..509)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     883..902
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      960..969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(966..971,972..992,993..2264)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     primer_bind     complement(2289..2306)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2295..2519
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2613..2632)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2614..2632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2639..2656)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2639..2655)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2648..2670)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2796..3251)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2933..2954)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3145..3164
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3278..3382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3383..3451,3452..4243)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3601..3620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4414..5002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4903..4922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
      361 gaaattaacc ctcactaaag ggaacaaaag ctggtaccgg gccccccctc gaggtcgacg
      421 gtatcgataa gcttgatatc gaattctacc gggtagggga ggcgcttttc ccaaggcagt
      481 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
      541 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
      601 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
      661 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
      721 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
      781 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
      841 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
      901 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg accatcgccg
      961 ccaccatggc tcctaagaag aagaggaagg tgatgagcca gttcgacatc ctgtgcaaga
     1021 ccccccccaa ggtgctggtg cggcagttcg tggagagatt cgagaggccc agcggcgaga
     1081 agatcgccag ctgtgccgcc gagctgacct acctgtgctg gatgatcacc cacaacggca
     1141 ccgccatcaa gagggccacc ttcatgagct acaacaccat catcagcaac agcctgagct
     1201 tcgacatcgt gaacaagagc ctgcagttca agtacaagac ccagaaggcc accatcctgg
     1261 aggccagcct gaagaagctg atccccgcct gggagttcac catcatccct tacaacggcc
     1321 agaagcacca gagcgacatc accgacatcg tgtccagcct gcagctgcag ttcgagagca
     1381 gcgaggaggc cgacaagggc aacagccaca gcaagaagat gctgaaggcc ctgctgtccg
     1441 agggcgagag catctgggag atcaccgaga agatcctgaa cagcttcgag tacaccagca
     1501 ggttcaccaa gaccaagacc ctgtaccagt tcctgttcct ggccacattc atcaactgcg
     1561 gcaggttcag cgacatcaag aacgtggacc ccaagagctt caagctggtg cagaacaagt
     1621 acctgggcgt gatcattcag tgcctggtga ccgagaccaa gacaagcgtg tccaggcaca
     1681 tctacttttt cagcgccaga ggcaggatcg accccctggt gtacctggac gagttcctga
     1741 ggaacagcga gcccgtgctg aagagagtga acaggaccgg caacagcagc agcaacaagc
     1801 aggagtacca gctgctgaag gacaacctgg tgcgcagcta caacaaggcc ctgaagaaga
     1861 acgcccccta ccccatcttc gctatcaaga acggccctaa gagccacatc ggcaggcacc
     1921 tgatgaccag ctttctgagc atgaagggcc tgaccgagct gacaaacgtg gtgggcaact
     1981 ggagcgacaa gagggcctcc gccgtggcca ggaccaccta cacccaccag atcaccgcca
     2041 tccccgacca ctacttcgcc ctggtgtcca ggtactacgc ctacgacccc atcagcaagg
     2101 agatgatcgc cctgaaggac gagaccaacc ccatcgagga gtggcagcac atcgagcagc
     2161 tgaagggcag cgccgagggc agcatcagat accccgcctg gaacggcatc atcagccagg
     2221 aggtgctgga ctacctgagc agctacatca acaggcggat ctgatgagat ctagagctcg
     2281 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2341 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2401 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2461 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2521 ttctgaggcg gaaagaacca gctggggctc gagatccact agttctagcc tcgaggctag
     2581 agcggccgcc accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac
     2641 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2701 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2761 cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta atattttgtt
     2821 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2881 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2941 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3001 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3061 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3121 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3181 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3241 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3301 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3361 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3421 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3481 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3541 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3601 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3661 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3721 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3781 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3841 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3901 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3961 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4021 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4081 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4141 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4201 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4261 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4321 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4381 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4441 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4501 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4561 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4621 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4681 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4741 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4801 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4861 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4921 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4981 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5041 ttgctggcct tttgctcaca tg
//
