LOCUS       sequence-247136-        6597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric TagBFP2 and dre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6597)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     2A plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6597)
  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            "Unpublished"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-dre
FEATURES             Location/Qualifiers
     source          1..6597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          138..1154
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1078..1100
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1162..1181
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1212..1922
                     /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             1944..1997
                     /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             3060..3080
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3098..3117)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3163..3218
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3217..3236)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3579..3595)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3579..3595)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3592..3614)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3603..3619
                     /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(3627..3633,3634..3651,3652..3657))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3672..3693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3752..3947
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3798..3933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3953..4087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3990..4009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4044..4063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4155..4172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4326..4914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4406..4425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5085..5876,5877..5945))
                     /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     5708..5727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5946..6050)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6081..6460
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
       61 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      121 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      181 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      241 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
      301 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
      361 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
      421 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
      481 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
      541 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
      601 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
      661 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
      721 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
      781 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
      841 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
      901 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
      961 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1021 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1081 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1141 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1201 aagaattcgc catggtgtct aagggcgaag agctgattaa ggagaacatg cacatgaagc
     1261 tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca
     1321 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
     1381 tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca
     1441 cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag
     1501 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
     1561 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga
     1621 tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg
     1681 gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa
     1741 acgccaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct
     1801 actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc
     1861 agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta
     1921 atggcggctc cggcggccgc tccgagggcc gcggctccct gctgacctgc ggcgacgtgg
     1981 aggagaaccc cggccccgcc cccggctcca ccatgggtgc tagcgagctg atcatctctg
     2041 gctcctctgg aggattcctg aggaacatcg gcaaggagta ccaggaggct gctgagaact
     2101 tcatgagatt catgaatgac cagggagcct acgcccctaa caccctgaga gacctgaggc
     2161 tggtgttcca ctcctgggct agatggtgcc acgctagaca gctggcctgg ttccctatct
     2221 ctcctgagat ggctagggag tacttccttc agctgcacga tgctgacctg gcctctacca
     2281 ccatcgacaa gcactacgcc atgctgaaca tgctgctgtc ccactgtggc ctgcctcctc
     2341 tgtctgatga caagtctgtg agcctggcca tgaggagaat ccggagagag gctgccaccg
     2401 agaagggaga gagaaccggc caggccatcc ctctgagatg ggatgacctg aagctgctgg
     2461 atgtgctgct gtctagatct gagagactgg tggacctgag gaatagggcc ttcctgtttg
     2521 tggcctacaa caccctgatg aggatgtctg agatctctag gatcagagtg ggagacctgg
     2581 accagaccgg agacaccgtg accctgcaca tctcccacac caagaccatc accaccgctg
     2641 ctggcctgga caaagtgctg tctaggagga ccaccgctgt gctgaatgac tggctggatg
     2701 tgtctggcct gagagagcac cctgacgctg tgctgttccc tcctatccac cggagcaaca
     2761 aggctaggat caccaccacc cctctgaccg cccctgccat ggagaagatt tttagcgatg
     2821 cctgggtgct gctgaacaag agggatgcca cccctaacaa gggccgctac cggacctgga
     2881 ccggccactc tgctagagtg ggagctgcca tcgacatggc tgagaagcaa gtgtccatgg
     2941 tggagatcat gcaggagggc acctggaaaa agcctgagac actgatgaga tacctgagga
     3001 ggggaggagt gtctgtggga gccaactcta ggctgatgga ctccgctagc ggcgccggtc
     3061 ctaagaagaa gaggaaagtg tgataaggcg cgccggatct ttttccctct gccaaaaatt
     3121 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     3181 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     3241 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     3301 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     3361 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     3421 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     3481 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     3541 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     3601 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3661 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3721 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     3781 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3841 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3901 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     3961 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4021 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4081 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4141 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4201 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4261 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4321 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4381 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4441 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4501 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4561 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4621 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4681 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4741 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4801 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4861 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4921 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4981 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5041 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5101 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5161 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5221 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5281 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5341 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5401 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5461 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5521 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5581 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5641 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5701 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5761 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5821 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5881 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5941 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6001 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6061 cgaaaagtgc cacctgggtc gacattgatt attgactagt tattaatagt aatcaattac
     6121 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6181 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6241 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6301 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6361 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6421 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     6481 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     6541 ttaattattt tgtgcagcga tgggggcggg gggggggggg ggggccaggc ggggcgg
//
