LOCUS       sequence-274442-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric TagBFP2 and iCre-ERT2 recombinase (PAA N-terminus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7496)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     2A creER optimization plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7496)
  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            "Unpublished"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 (PAA)
FEATURES             Location/Qualifiers
     source          1..7496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        11..390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        392..669
                     /label=chicken beta-actin promoter
     intron          670..1686
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1610..1632
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1744..2454
                     /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             2476..2529
                     /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             3571..4506
                     /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             3619..4512
                     /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(4524..4543)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4589..4644
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4643..4662)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5005..5021)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5005..5021)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5018..5040)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5029..5045
                     /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(5053..5059,5060..5077,5078..5083))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5098..5119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5178..5373
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5224..5359
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5379..5513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5416..5435)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5470..5489
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5581..5598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5752..6340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5832..5851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6511..7302,7303..7371))
                     /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     7134..7153
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7372..7476)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgggtc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
      421 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
      481 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc cggggggggg gggggggggg
      541 gggggggggg gsggggsgrg gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
      601 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
      661 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
      721 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
      781 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg
      841 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg
      901 ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc
      961 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc
     1021 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg
     1081 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc
     1141 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1201 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1261 ggcggggcgg ggccgcctcg ggccggggrg ggctcggggg aggggcgcgg cggcccccgg
     1321 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
     1381 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
     1441 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
     1501 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
     1561 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     1621 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     1741 gccatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg
     1801 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
     1861 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     1921 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
     1981 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
     2041 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     2101 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     2161 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     2221 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag
     2281 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     2341 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     2401 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taatggcggc
     2461 tccggcggcc gctccgaggg ccgcggcagc ctgctgacat gcggcgacgt ggaagagaac
     2521 cctggccctg ccgctaacct gctgactgtg caccaaaacc tgcctgccct ccctgtggat
     2581 gccacctctg atgaagtcag gaagaacctg atggacatgt tcagggacag gcaggccttc
     2641 tctgaacaca cctggaagat gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag
     2701 ctgaacaaca ggaaatggtt ccctgctgaa cctgaggatg tgagggacta cctcctgtac
     2761 ctgcaagcca gaggcctggc tgtgaagacc atccaacagc acctgggcca gctcaacatg
     2821 ctgcacagga gatctggcct gcctcgccct tctgactcca atgctgtgtc cctggtgatg
     2881 aggagaatca gaaaggagaa tgtggatgct ggggagagag ccaagcaggc cctggccttt
     2941 gaacgcactg actttgacca agtcagatcc ctgatggaga actctgacag atgccaggac
     3001 atcaggaacc tggccttcct gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt
     3061 gccagaatca gagtgaagga catctcccgc accgatggtg ggagaatgct gatccacatt
     3121 ggcaggacca agaccctggt gtccacagct ggtgtggaga aggccctgtc cctgggggtt
     3181 accaagctgg tggagagatg gatctctgtg tctggtgtgg ctgatgaccc caacaactac
     3241 ctgttctgcc gggtcagaaa gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc
     3301 acccgggccc tggaagggat ctttgaggcc acccaccgcc tgatctatgg tgccaaggat
     3361 gactctgggc agagatacct ggcctggtct ggccactctg ccagagtggg tgctgccagg
     3421 gacatggcca gggctggtgt gtccatccct gaaatcatgc aggctggtgg ctggaccaat
     3481 gtgaacattg tgatgaacta catcagaaac ctggactctg agactggggc catggtgagg
     3541 ctgctagagg atggggacct cgagccatct gctggagaca tgagagctgc caacctttgg
     3601 ccaagcccgc tcatgatcaa acgctctaag aagaacagcc tggccttgtc cctgacggcc
     3661 gaccagatgg tcagtgcctt gttggatgct gagcccccca tactctattc cgagtatgat
     3721 cctaccagac ccttcagtga agcttcgatg atgggcttac tgaccaacct ggcagacagg
     3781 gagctggttc acatgatcaa ctgggcgaag agggtgccag gctttgtgga tttgaccctc
     3841 catgatcagg tccaccttct agaatgtgcc tggctagaga tcctgatgat tggtctcgtc
     3901 tggcgctcca tggagcaccc agtgaagcta ctgtttgctc ctaacttgct cttggacagg
     3961 aaccagggaa aatgtgtaga gggcatggtg gagatcttcg acatgctgct ggctacatca
     4021 tctcggttcc gcatgatgaa tctgcaggga gaggagtttg tgtgcctcaa atctattatt
     4081 ttgcttaatt ctggagtgta cacatttctg tccagcaccc tgaagtctct ggaagagaag
     4141 gaccatatcc accgagtcct ggacaagatc acagacactt tgatccacct gatggccaag
     4201 gcaggcctga ccctgcagca gcagcaccag cggctggccc agctcctcct catcctctcc
     4261 cacatcaggc acatgagtaa caaaggcatg gagcatctgt acagcatgaa gtgcaagaac
     4321 gtggtgcccc tctatgacct gctgctggag gcggcggacg cccaccgcct acatgcgccc
     4381 actagccgtg gaggggcatc cgtggaggag acggaccaaa gccacttggc cactgcgggc
     4441 tctacttcat cgcattcctt gcaaaagtat tacatcacgg gggaggcaga gggtttccct
     4501 gccacagctt gaggcgcgcc ggatcttttt ccctctgcca aaaattatgg ggacatcatg
     4561 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     4621 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     4681 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     4741 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     4801 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     4861 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     4921 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     4981 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5041 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5101 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5161 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5221 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5281 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5341 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     5401 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5461 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     5521 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5581 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5641 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5701 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5761 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5821 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5881 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5941 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6001 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6061 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6121 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6181 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6241 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6301 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6361 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6421 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6481 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6541 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6601 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6661 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6721 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6781 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6841 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6901 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6961 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7021 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7081 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7141 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7201 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7261 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7321 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7381 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7441 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgc
//
