LOCUS       sequence-247964-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric TagBFP2 and iCre-ERT2 recombinase (PVV 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  JOURNAL   Unpublished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            "Unpublished"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 (PVV)
FEATURES             Location/Qualifiers
     source          1..7496
                     /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             3561..4496
                     /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             3609..4502
                     /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(4514..4533)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4579..4634
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4633..4652)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4995..5011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4995..5011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5008..5030)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5019..5035
                     /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(5043..5049,5050..5067,5068..5073))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5088..5109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5168..5363
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5214..5349
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5369..5503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5406..5425)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5460..5479
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5571..5588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5742..6330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5822..5841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6501..7292,7293..7361))
                     /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     7124..7143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7362..7466)
                     /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 tggtgaacct gctgactgtg caccaaaacc tgcctgccct ccctgtggat gccacctctg
     2581 atgaagtcag gaagaacctg atggacatgt tcagggacag gcaggccttc tctgaacaca
     2641 cctggaagat gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag ctgaacaaca
     2701 ggaaatggtt ccctgctgaa cctgaggatg tgagggacta cctcctgtac ctgcaagcca
     2761 gaggcctggc tgtgaagacc atccaacagc acctgggcca gctcaacatg ctgcacagga
     2821 gatctggcct gcctcgccct tctgactcca atgctgtgtc cctggtgatg aggagaatca
     2881 gaaaggagaa tgtggatgct ggggagagag ccaagcaggc cctggccttt gaacgcactg
     2941 actttgacca agtcagatcc ctgatggaga actctgacag atgccaggac atcaggaacc
     3001 tggccttcct gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt gccagaatca
     3061 gagtgaagga catctcccgc accgatggtg ggagaatgct gatccacatt ggcaggacca
     3121 agaccctggt gtccacagct ggtgtggaga aggccctgtc cctgggggtt accaagctgg
     3181 tggagagatg gatctctgtg tctggtgtgg ctgatgaccc caacaactac ctgttctgcc
     3241 gggtcagaaa gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc acccgggccc
     3301 tggaagggat ctttgaggcc acccaccgcc tgatctatgg tgccaaggat gactctgggc
     3361 agagatacct ggcctggtct ggccactctg ccagagtggg tgctgccagg gacatggcca
     3421 gggctggtgt gtccatccct gaaatcatgc aggctggtgg ctggaccaat gtgaacattg
     3481 tgatgaacta catcagaaac ctggactctg agactggggc catggtgagg ctgctagagg
     3541 atggggacct cgagccatct gctggagaca tgagagctgc caacctttgg ccaagcccgc
     3601 tcatgatcaa acgctctaag aagaacagcc tggccttgtc cctgacggcc gaccagatgg
     3661 tcagtgcctt gttggatgct gagcccccca tactctattc cgagtatgat cctaccagac
     3721 ccttcagtga agcttcgatg atgggcttac tgaccaacct ggcagacagg gagctggttc
     3781 acatgatcaa ctgggcgaag agggtgccag gctttgtgga tttgaccctc catgatcagg
     3841 tccaccttct agaatgtgcc tggctagaga tcctgatgat tggtctcgtc tggcgctcca
     3901 tggagcaccc agtgaagcta ctgtttgctc ctaacttgct cttggacagg aaccagggaa
     3961 aatgtgtaga gggcatggtg gagatcttcg acatgctgct ggctacatca tctcggttcc
     4021 gcatgatgaa tctgcaggga gaggagtttg tgtgcctcaa atctattatt ttgcttaatt
     4081 ctggagtgta cacatttctg tccagcaccc tgaagtctct ggaagagaag gaccatatcc
     4141 accgagtcct ggacaagatc acagacactt tgatccacct gatggccaag gcaggcctga
     4201 ccctgcagca gcagcaccag cggctggccc agctcctcct catcctctcc cacatcaggc
     4261 acatgagtaa caaaggcatg gagcatctgt acagcatgaa gtgcaagaac gtggtgcccc
     4321 tctatgacct gctgctggag gcggcggacg cccaccgcct acatgcgccc actagccgtg
     4381 gaggggcatc cgtggaggag acggaccaaa gccacttggc cactgcgggc tctacttcat
     4441 cgcattcctt gcaaaagtat tacatcacgg gggaggcaga gggtttccct gccacagctt
     4501 gaggcgcgcc ggatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     4561 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4621 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4681 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     4741 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4801 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4861 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     4921 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc
     4981 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5041 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5101 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5161 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5221 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5281 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5341 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
     5401 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5461 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
     5521 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5581 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5641 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5701 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5761 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5821 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5881 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5941 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6001 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6061 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6121 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6181 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6241 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6301 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6361 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6421 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6481 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6541 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6601 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6661 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6721 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6781 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6841 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6901 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6961 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7021 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7081 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7141 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7201 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7261 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7321 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7381 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7441 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtc
//
