LOCUS       sequence-274446-        7499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric TagBFP2 and iCre-ERT2 recombinase (PAAA N-terminus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7499)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     2A creER optimization plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7499)
  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            "Unpublished"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 (PAAA)
FEATURES             Location/Qualifiers
     source          1..7499
                     /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             3574..4509
                     /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             3622..4515
                     /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(4527..4546)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4592..4647
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4646..4665)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5008..5024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5008..5024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5021..5043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5032..5048
                     /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(5056..5062,5063..5080,5081..5086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5101..5122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5181..5376
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5227..5362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5382..5516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5419..5438)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5473..5492
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5584..5601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5755..6343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5835..5854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6514..7305,7306..7374))
                     /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     7137..7156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7375..7479)
                     /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 rggggcgcgg 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 ccgctgcaaa cctgctgact gtgcaccaaa acctgcctgc cctccctgtg
     2581 gatgccacct ctgatgaagt caggaagaac ctgatggaca tgttcaggga caggcaggcc
     2641 ttctctgaac acacctggaa gatgctcctg tctgtgtgca gatcctgggc tgcctggtgc
     2701 aagctgaaca acaggaaatg gttccctgct gaacctgagg atgtgaggga ctacctcctg
     2761 tacctgcaag ccagaggcct ggctgtgaag accatccaac agcacctggg ccagctcaac
     2821 atgctgcaca ggagatctgg cctgcctcgc ccttctgact ccaatgctgt gtccctggtg
     2881 atgaggagaa tcagaaagga gaatgtggat gctggggaga gagccaagca ggccctggcc
     2941 tttgaacgca ctgactttga ccaagtcaga tccctgatgg agaactctga cagatgccag
     3001 gacatcagga acctggcctt cctgggcatt gcctacaaca ccctgctgcg cattgccgaa
     3061 attgccagaa tcagagtgaa ggacatctcc cgcaccgatg gtgggagaat gctgatccac
     3121 attggcagga ccaagaccct ggtgtccaca gctggtgtgg agaaggccct gtccctgggg
     3181 gttaccaagc tggtggagag atggatctct gtgtctggtg tggctgatga ccccaacaac
     3241 tacctgttct gccgggtcag aaagaatggt gtggctgccc cttctgccac ctcccaactg
     3301 tccacccggg ccctggaagg gatctttgag gccacccacc gcctgatcta tggtgccaag
     3361 gatgactctg ggcagagata cctggcctgg tctggccact ctgccagagt gggtgctgcc
     3421 agggacatgg ccagggctgg tgtgtccatc cctgaaatca tgcaggctgg tggctggacc
     3481 aatgtgaaca ttgtgatgaa ctacatcaga aacctggact ctgagactgg ggccatggtg
     3541 aggctgctag aggatgggga cctcgagcca tctgctggag acatgagagc tgccaacctt
     3601 tggccaagcc cgctcatgat caaacgctct aagaagaaca gcctggcctt gtccctgacg
     3661 gccgaccaga tggtcagtgc cttgttggat gctgagcccc ccatactcta ttccgagtat
     3721 gatcctacca gacccttcag tgaagcttcg atgatgggct tactgaccaa cctggcagac
     3781 agggagctgg ttcacatgat caactgggcg aagagggtgc caggctttgt ggatttgacc
     3841 ctccatgatc aggtccacct tctagaatgt gcctggctag agatcctgat gattggtctc
     3901 gtctggcgct ccatggagca cccagtgaag ctactgtttg ctcctaactt gctcttggac
     3961 aggaaccagg gaaaatgtgt agagggcatg gtggagatct tcgacatgct gctggctaca
     4021 tcatctcggt tccgcatgat gaatctgcag ggagaggagt ttgtgtgcct caaatctatt
     4081 attttgctta attctggagt gtacacattt ctgtccagca ccctgaagtc tctggaagag
     4141 aaggaccata tccaccgagt cctggacaag atcacagaca ctttgatcca cctgatggcc
     4201 aaggcaggcc tgaccctgca gcagcagcac cagcggctgg cccagctcct cctcatcctc
     4261 tcccacatca ggcacatgag taacaaaggc atggagcatc tgtacagcat gaagtgcaag
     4321 aacgtggtgc ccctctatga cctgctgctg gaggcggcgg acgcccaccg cctacatgcg
     4381 cccactagcc gtggaggggc atccgtggag gagacggacc aaagccactt ggccactgcg
     4441 ggctctactt catcgcattc cttgcaaaag tattacatca cgggggaggc agagggtttc
     4501 cctgccacag cttgaggcgc gccggatctt tttccctctg ccaaaaatta tggggacatc
     4561 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     4621 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     4681 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     4741 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     4801 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     4861 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     4921 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     4981 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5041 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5101 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5161 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5221 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5281 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5341 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     5401 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5461 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     5521 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5581 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5641 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5941 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6481 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6541 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6601 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6661 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6721 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6781 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6841 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6901 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6961 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7021 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7081 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7141 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7201 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7261 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7321 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7381 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7441 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgc
//
