LOCUS       sequence-247966-        7493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric TagBFP2 and iCre-ERT2 recombinase (PA N-terminus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7493)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     2A creER optimization plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7493)
  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            "Unpublished"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-iCreERT2 (PA)
FEATURES             Location/Qualifiers
     source          1..7493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1734..2444
                     /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             2466..2519
                     /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             3558..4493
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             3606..4499
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     primer_bind     complement(4511..4530)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4576..4631
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4630..4649)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4992..5008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4992..5008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5005..5027)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5016..5032
                     /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(5040..5046,5047..5064,5065..5070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5085..5106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5165..5360
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5211..5346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5366..5500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5403..5422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5457..5476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5568..5585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5739..6327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5819..5838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6498..7289,7290..7358))
                     /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     7121..7140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7359..7463)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc ggggggcggg
      541 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc gccatggtgt
     1741 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     1801 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     1861 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     1921 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
     1981 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     2041 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     2101 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
     2161 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
     2221 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata
     2281 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
     2341 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
     2401 ccagatactg cgacctccct agcaaactgg ggcacaagct taatggcggc tccggcggcc
     2461 gctccgaggg ccgcggcagc ctgctgacat gcggcgacgt ggaagagaac cctggccctg
     2521 ccaacctgct gactgtgcac caaaacctgc ctgccctccc tgtggatgcc acctctgatg
     2581 aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct gaacacacct
     2641 ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg aacaacagga
     2701 aatggttccc tgctgaacct gaggatgtga gggactacct cctgtacctg caagccagag
     2761 gcctggctgt gaagaccatc caacagcacc tgggccagct caacatgctg cacaggagat
     2821 ctggcctgcc tcgcccttct gactccaatg ctgtgtccct ggtgatgagg agaatcagaa
     2881 aggagaatgt ggatgctggg gagagagcca agcaggccct ggcctttgaa cgcactgact
     2941 ttgaccaagt cagatccctg atggagaact ctgacagatg ccaggacatc aggaacctgg
     3001 ccttcctggg cattgcctac aacaccctgc tgcgcattgc cgaaattgcc agaatcagag
     3061 tgaaggacat ctcccgcacc gatggtggga gaatgctgat ccacattggc aggaccaaga
     3121 ccctggtgtc cacagctggt gtggagaagg ccctgtccct gggggttacc aagctggtgg
     3181 agagatggat ctctgtgtct ggtgtggctg atgaccccaa caactacctg ttctgccggg
     3241 tcagaaagaa tggtgtggct gccccttctg ccacctccca actgtccacc cgggccctgg
     3301 aagggatctt tgaggccacc caccgcctga tctatggtgc caaggatgac tctgggcaga
     3361 gatacctggc ctggtctggc cactctgcca gagtgggtgc tgccagggac atggccaggg
     3421 ctggtgtgtc catccctgaa atcatgcagg ctggtggctg gaccaatgtg aacattgtga
     3481 tgaactacat cagaaacctg gactctgaga ctggggccat ggtgaggctg ctagaggatg
     3541 gggacctcga gccatctgct ggagacatga gagctgccaa cctttggcca agcccgctca
     3601 tgatcaaacg ctctaagaag aacagcctgg ccttgtccct gacggccgac cagatggtca
     3661 gtgccttgtt ggatgctgag ccccccatac tctattccga gtatgatcct accagaccct
     3721 tcagtgaagc ttcgatgatg ggcttactga ccaacctggc agacagggag ctggttcaca
     3781 tgatcaactg ggcgaagagg gtgccaggct ttgtggattt gaccctccat gatcaggtcc
     3841 accttctaga atgtgcctgg ctagagatcc tgatgattgg tctcgtctgg cgctccatgg
     3901 agcacccagt gaagctactg tttgctccta acttgctctt ggacaggaac cagggaaaat
     3961 gtgtagaggg catggtggag atcttcgaca tgctgctggc tacatcatct cggttccgca
     4021 tgatgaatct gcagggagag gagtttgtgt gcctcaaatc tattattttg cttaattctg
     4081 gagtgtacac atttctgtcc agcaccctga agtctctgga agagaaggac catatccacc
     4141 gagtcctgga caagatcaca gacactttga tccacctgat ggccaaggca ggcctgaccc
     4201 tgcagcagca gcaccagcgg ctggcccagc tcctcctcat cctctcccac atcaggcaca
     4261 tgagtaacaa aggcatggag catctgtaca gcatgaagtg caagaacgtg gtgcccctct
     4321 atgacctgct gctggaggcg gcggacgccc accgcctaca tgcgcccact agccgtggag
     4381 gggcatccgt ggaggagacg gaccaaagcc acttggccac tgcgggctct acttcatcgc
     4441 attccttgca aaagtattac atcacggggg aggcagaggg tttccctgcc acagcttgag
     4501 gcgcgccgga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     4561 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     4621 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     4681 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     4741 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     4801 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     4861 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     4921 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg
     4981 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5041 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5101 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     5161 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5221 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5281 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5341 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     5401 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5461 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     5521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtc
//
