LOCUS       sequence-247146-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric TagBFP2 and FLP recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     2A plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6827)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCAGGS-mTagBFP2-T2A-FLPo
FEATURES             Location/Qualifiers
     source          1..6827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          133..1149
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1073..1095
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1157..1176
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1207..1917
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             1939..1992
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(2008..2013,2014..2034,2035..3306)
                     /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(3321..3340)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3386..3441
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3440..3459)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3802..3818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3802..3818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3815..3837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3826..3842
                     /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)."
     promoter        complement(join(3850..3856,3857..3874,3875..3880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3895..3916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3975..4170
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4021..4156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4176..4310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4213..4232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4267..4286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4378..4395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4549..5137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4629..4648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5308..6099,6100..6168))
                     /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     5931..5950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6169..6273)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6304..6683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6685..6827,1..132)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
       61 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      121 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      181 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      241 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      301 tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg
      361 gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc
      421 ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg
      481 agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc
      541 gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa
      601 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
      661 cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg
      721 ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc
      781 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
      841 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
      901 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
      961 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     1021 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1081 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     1141 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     1201 ttcgccatgg tgtctaaggg cgaagagctg attaaggaga acatgcacat gaagctgtac
     1261 atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc
     1321 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
     1381 ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag
     1441 ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc
     1501 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
     1561 ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag
     1621 aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg
     1681 gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacgcc
     1741 aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat
     1801 gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac
     1861 gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaatggc
     1921 ggctccggcg gccgctccga gggccgcggc tccctgctga cctgcggcga cgtggaggag
     1981 aaccccggcc ccgcccccgg ctccaccatg gctcctaaga agaagaggaa ggtgatgagc
     2041 cagttcgaca tcctgtgcaa gacccccccc aaggtgctgg tgcggcagtt cgtggagaga
     2101 ttcgagaggc ccagcggcga gaagatcgcc agctgtgccg ccgagctgac ctacctgtgc
     2161 tggatgatca cccacaacgg caccgccatc aagagggcca ccttcatgag ctacaacacc
     2221 atcatcagca acagcctgag cttcgacatc gtgaacaaga gcctgcagtt caagtacaag
     2281 acccagaagg ccaccatcct ggaggccagc ctgaagaagc tgatccccgc ctgggagttc
     2341 accatcatcc cttacaacgg ccagaagcac cagagcgaca tcaccgacat cgtgtccagc
     2401 ctgcagctgc agttcgagag cagcgaggag gccgacaagg gcaacagcca cagcaagaag
     2461 atgctgaagg ccctgctgtc cgagggcgag agcatctggg agatcaccga gaagatcctg
     2521 aacagcttcg agtacaccag caggttcacc aagaccaaga ccctgtacca gttcctgttc
     2581 ctggccacat tcatcaactg cggcaggttc agcgacatca agaacgtgga ccccaagagc
     2641 ttcaagctgg tgcagaacaa gtacctgggc gtgatcattc agtgcctggt gaccgagacc
     2701 aagacaagcg tgtccaggca catctacttt ttcagcgcca gaggcaggat cgaccccctg
     2761 gtgtacctgg acgagttcct gaggaacagc gagcccgtgc tgaagagagt gaacaggacc
     2821 ggcaacagca gcagcaacaa gcaggagtac cagctgctga aggacaacct ggtgcgcagc
     2881 tacaacaagg ccctgaagaa gaacgccccc taccccatct tcgctatcaa gaacggccct
     2941 aagagccaca tcggcaggca cctgatgacc agctttctga gcatgaaggg cctgaccgag
     3001 ctgacaaacg tggtgggcaa ctggagcgac aagagggcct ccgccgtggc caggaccacc
     3061 tacacccacc agatcaccgc catccccgac cactacttcg ccctggtgtc caggtactac
     3121 gcctacgacc ccatcagcaa ggagatgatc gccctgaagg acgagaccaa ccccatcgag
     3181 gagtggcagc acatcgagca gctgaagggc agcgccgagg gcagcatcag ataccccgcc
     3241 tggaacggca tcatcagcca ggaggtgctg gactacctga gcagctacat caacaggcgg
     3301 atctgataag gcgcgccgga tctttttccc tctgccaaaa attatgggga catcatgaag
     3361 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     3421 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     3481 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     3541 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     3601 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     3661 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     3721 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     3781 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3841 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3961 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4021 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4081 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4141 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     4201 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4261 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     4321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4981 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5041 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5281 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5341 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5401 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5461 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5521 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5581 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5641 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5701 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5761 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5821 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5881 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5941 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6001 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6061 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6121 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6181 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6241 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     6301 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     6361 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6421 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     6481 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     6541 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6601 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6661 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     6721 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     6781 cgatgggggc gggggggggg gggggsgcgc caggcggggc ggggcgg
//
