LOCUS       sequence-304997-        8142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  episomal expression of FlpO and Cre recombinases, self excising.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8142)
  AUTHORS   Kim GB, Rincon Fernandez Pacheco D, Saxon D, Yang A, Sabet S, 
            Dutra-Clarke M, Levy R, Watkins A, Park H, Abbasi Akhtar A, Linesch 
            PW, Kobritz N, Chandra SS, Grausam K, Ayala-Sarmiento A, Molina J, 
            Sedivakova K, Hoang K, Tsyporin J, Gareau DS, Filbin MG, Bannykh S, 
            Santiskulvong C, Wang Y, Tang J, Suva ML, Chen B, Danielpour M, 
            Breunig JJ
  TITLE     Rapid Generation of Somatic Mouse Mosaics with Locus-Specific, 
            Stably Integrated Transgenic Elements.
  JOURNAL   Cell. 2019 Sep 19;179(1):251-267.e24. doi: 
            10.1016/j.cell.2019.08.013.
  PUBMED    31539496
REFERENCE   2  (bases 1 to 8142)
  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"; doi: 
            "10.1016/j.cell.2019.08"; journalName: "Cell"; date: "2019-09-19- 
            19"; volume: "179"; issue: "1"; pages: "251-267.e24"
COMMENT     Sequence Label: pCag SE (Self-excising) FlpO-2A-Cre EV
FEATURES             Location/Qualifiers
     source          1..8142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          160..1176
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1100..1122
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1184..1203
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1289..1518
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1501..1520
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2971..3024
                     /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             3025..4053
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     polyA_signal    4224..4345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4235..4254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4289..4308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4560..4580)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4560..4578)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4599..4615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4599..4615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4612..4634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4623..4639
                     /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(4647..4653,4654..4671,4672..4677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4692..4713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4830..4847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5001..5589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5081..5100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5760..6551,6552..6620))
                     /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     6383..6402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6621..6725)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6751..7206)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6888..6909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7100..7119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7333..7355
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7347..7364
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7348..7364
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7374..7392
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        7645..8024
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8026..8142,1..159)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggggcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
       61 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      121 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      181 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      241 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      301 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      361 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg tgtggggagc
      421 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
      481 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
      541 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
      601 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
      661 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
      721 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
      781 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
      841 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
      901 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
      961 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1021 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1081 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1141 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1201 gtgctgtctc atcattttgg caaatacgac tcactatagg gagacccaag cttctagaat
     1261 tcgctgtctg cgagggccag ctgttggggt gagtactccc tctcaaaagc gggcatgact
     1321 tctgcgctaa gattgtcagt ttccaaaaac gaggaggatt tgatattcac ctggcccgcg
     1381 gtgatgcctt tgagggtggc cgcgtccatc tggtcagaaa agacaatctt tttgttgtca
     1441 agcttgaggt gtggcaggct tgagatctgg ccatacactt gagtgacaat gacatccact
     1501 ttgcctttct ctccacaggt gtccactccc aggtccaact gcagcccaag ctaggaagtt
     1561 cctattcttc aaaaggtata ggaacttcgc cgccaccatg agccagttcg acatcctgtg
     1621 caagaccccc cccaaggtgc tggtgcggca gttcgtggag agattcgaga ggcccagcgg
     1681 cgagaagatc gccagctgtg ccgccgagct gacctacctg tgctggatga tcacccacaa
     1741 cggcaccgcc atcaagaggg ccaccttcat gagctacaac accatcatca gcaacagcct
     1801 gagcttcgac atcgtgaaca agagcctgca gttcaagtac aagacccaga aggccaccat
     1861 cctggaggcc agcctgaaga agctgatccc cgcctgggag ttcaccatca tcccttacaa
     1921 cggccagaag caccagagcg acatcaccga catcgtgtcc agcctgcagc tgcagttcga
     1981 gagcagcgag gaggccgaca agggcaacag ccacagcaag aagatgctga aggccctgct
     2041 gtccgagggc gagagcatct gggagatcac cgagaagatc ctgaacagct tcgagtacac
     2101 cagcaggttc accaagacca agaccctgta ccagttcctg ttcctggcca cattcatcaa
     2161 ctgcggcagg ttcagcgaca tcaagaacgt ggaccccaag agcttcaagc tggtgcagaa
     2221 caagtacctg ggcgtgatca ttcagtgcct ggtgaccgag accaagacaa gcgtgtccag
     2281 gcacatctac tttttcagcg ccagaggcag gatcgacccc ctggtgtacc tggacgagtt
     2341 cctgaggaac agcgagcccg tgctgaagag agtgaacagg accggcaaca gcagcagcaa
     2401 caagcaggag taccagctgc tgaaggacaa cctggtgcgc agctacaaca aggccctgaa
     2461 gaagaacgcc ccctacccca tcttcgctat caagaacggc cctaagagcc acatcggcag
     2521 gcacctgatg accagctttc tgagcatgaa gggcctgacc gagctgacaa acgtggtggg
     2581 caactggagc gacaagaggg cctccgccgt ggccaggacc acctacaccc accagatcac
     2641 cgccatcccc gaccactact tcgccctggt gtccaggtaa gtagagggct gggctgggct
     2701 gtggggggtg tggggtgcgg gactgggcag tctgggagtc cctctcacca cttttcttac
     2761 ctttctaggt actacgccta cgaccccatc agcaaggaga tgatcgccct gaaggacgag
     2821 accaacccca tcgaggagtg gcagcacatc gagcagctga agggcagcgc cgagggcagc
     2881 atcagatacc ccgcctggaa cggcatcatc agccaggagg tgctggacta cctgagcagc
     2941 tacatcaaca ggcggatctc tagaagagcc gagggcaggg gaagtcttct aacatgcggg
     3001 gacgtggagg aaaatcccgg gccctccaat ttactgaccg tacaccaaaa tttgcctgca
     3061 ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat
     3121 cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg
     3181 gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat
     3241 tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc
     3301 cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt
     3361 tcactggtta tgcggcgcat tcgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag
     3421 gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat
     3481 cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt
     3541 atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg
     3601 ttaatccata ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt
     3661 agcctggggg taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat
     3721 ccgaataact acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc
     3781 agccagctat caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac
     3841 ggcgctaagg atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc
     3901 ggagccgcgc gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt
     3961 ggctggacca atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg
     4021 gcaatggtgc gcctgctgga agatggcgat taggtacgca tttaaggaag ttcctattct
     4081 tcaaaaggta taggaacttc cataaacacg cactatgccg ttcttctcat gtatatatat
     4141 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgggtaccg
     4201 agctcctcga ggatcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4261 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4321 cattataagc tgcaataaac aagttatcct cgaggagctc atgagcgctt gtttcggcgt
     4381 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
     4441 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     4501 ggagtcgcat aagggagagc gtcgacctcg agggggggcc cggtacccag cttttgttcc
     4561 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     4621 aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc
     4681 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4741 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5161 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5401 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5701 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5761 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5821 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5881 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5941 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6001 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6061 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6121 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6181 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6241 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6301 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6361 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6421 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6481 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6541 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6601 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6661 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6721 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     6781 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     6841 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     6901 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     6961 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     7021 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     7081 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     7141 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     7201 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     7261 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     7321 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     7381 actcactata gggcgaattg gagctccacc gcggtctttc ggacttttga aagtgatggt
     7441 ggtgggggaa ggattcgaac cttcgaagtc gatgacggca gatttagagt ctgctccctg
     7501 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7561 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     7621 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     7681 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     7741 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     7801 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     7861 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     7921 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     7981 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     8041 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     8101 ttttttaatt attttgtgca gcgatggggg cggggggggg gg
//
