LOCUS       sequence-250194-        7971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  episomal expression of FlpO and Cre recombinases.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  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 7971)
  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 FlpO-2A-Cre EV
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        432..707
                     /label=chicken beta-actin promoter
     intron          708..1720
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1644..1666
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1728..1747
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1833..2062
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2045..2064
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3383..3436
                     /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             3437..4465
                     /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    4608..4729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4619..4638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4673..4692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4944..4964)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4944..4962)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4983..4999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4983..4999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4996..5018)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5007..5023
                     /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(5031..5037,5038..5055,5056..5061))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5076..5097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5214..5231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5385..5966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5465..5484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6137..6928,6929..6997))
                     /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     6760..6779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6998..7102)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7127..7582)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7264..7285)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7476..7495
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7709..7731
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7723..7740
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7724..7740
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7750..7768
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      481 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      541 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
      601 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
      661 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgt
      721 tgccttcgcc ccgtgccccg ctccgcgccg cctcgcgccg cccgccccgg ctctgactga
      781 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      841 gcttggttta atgacggctc gtttcttttc tgtggctgcg tgaaagcctt aaagggctcc
      901 gggagggccc tttgtgcggg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
      961 gggagcgccg cgtgcggccc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
     1021 ggctttgtgc gctccgcgtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
     1081 cgggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
     1141 gggggtgtgg gcgcggcggt cgggctgtaa cccccccctg cacccccctc cccgagttgc
     1201 tgagcacggc ccggcttcgg gtgcggggct ccgtgcgggg cgtggcgcgg ggctcgccgt
     1261 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     1321 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     1381 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     1441 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     1501 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     1561 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     1621 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1681 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1741 tattgtgctg tctcatcatt ttggcaaata cgactcacta tagggagacc caagcttcta
     1801 gaattcgctg tctgcgaggg ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     1861 gacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
     1921 cgcggtgatg cctttgaggg tggccgcgtc catctggtca gaaaagacaa tctttttgtt
     1981 gtcaagcttg aggtgtggca ggcttgagat ctggccatac acttgagtga caatgacatc
     2041 cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcagcc caagctagac
     2101 catgagccag ttcgacatcc tgtgcaagac cccccccaag gtgctggtgc ggcagttcgt
     2161 ggagagattc gagaggccca gcggcgagaa gatcgccagc tgtgccgccg agctgaccta
     2221 cctgtgctgg atgatcaccc acaacggcac cgccatcaag agggccacct tcatgagcta
     2281 caacaccatc atcagcaaca gcctgagctt cgacatcgtg aacaagagcc tgcagttcaa
     2341 gtacaagacc cagaaggcca ccatcctgga ggccagcctg aagaagctga tccccgcctg
     2401 ggagttcacc atcatccctt acaacggcca gaagcaccag agcgacatca ccgacatcgt
     2461 gtccagcctg cagctgcagt tcgagagcag cgaggaggcc gacaagggca acagccacag
     2521 caagaagatg ctgaaggccc tgctgtccga gggcgagagc atctgggaga tcaccgagaa
     2581 gatcctgaac agcttcgagt acaccagcag gttcaccaag accaagaccc tgtaccagtt
     2641 cctgttcctg gccacattca tcaactgcgg caggttcagc gacatcaaga acgtggaccc
     2701 caagagcttc aagctggtgc agaacaagta cctgggcgtg atcattcagt gcctggtgac
     2761 cgagaccaag acaagcgtgt ccaggcacat ctactttttc agcgccagag gcaggatcga
     2821 ccccctggtg tacctggacg agttcctgag gaacagcgag cccgtgctga agagagtgaa
     2881 caggaccggc aacagcagca gcaacaagca ggagtaccag ctgctgaagg acaacctggt
     2941 gcgcagctac aacaaggccc tgaagaagaa cgccccctac cccatcttcg ctatcaagaa
     3001 cggccctaag agccacatcg gcaggcacct gatgaccagc tttctgagca tgaagggcct
     3061 gaccgagctg acaaacgtgg tgggcaactg gagcgacaag agggcctccg ccgtggccag
     3121 gaccacctac acccaccaga tcaccgccat ccccgaccac tacttcgccc tggtgtccag
     3181 gtactacgcc tacgacccca tcagcaagga gatgatcgcc ctgaaggacg agaccaaccc
     3241 catcgaggag tggcagcaca tcgagcagct gaagggcagc gccgagggca gcatcagata
     3301 ccccgcctgg aacggcatca tcagccagga ggtgctggac tacctgagca gctacatcaa
     3361 caggcggatc tctagaagag ccgagggcag gggaagtctt ctaacatgcg gggacgtgga
     3421 ggaaaatccc gggccctcca atttactgac cgtacaccaa aatttgcctg cattaccggt
     3481 cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg atcgccaggc
     3541 gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg
     3601 caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg attatcttct
     3661 atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg gccagctaaa
     3721 catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg tttcactggt
     3781 tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc
     3841 gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg atcgctgcca
     3901 ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac gtatagccga
     3961 aattgccagg atcagggtta aagatatctc acgtactgac ggtgggagaa tgttaatcca
     4021 tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac ttagcctggg
     4081 ggtaactaaa ctggtcgagc gatggatttc cgtctctggt gtagctgatg atccgaataa
     4141 ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct
     4201 atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt acggcgctaa
     4261 ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg tcggagccgc
     4321 gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg gtggctggac
     4381 caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag gggcaatggt
     4441 gcgcctgctg gaagatggcg attaaggatc cgtacgcatt taagcataaa cacgcactat
     4501 gccgttcttc tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac
     4561 agtgagctgt atgtgcgggt accgagctcc tcgaggatca gacatgataa gatacattga
     4621 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4681 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta tcctcgagga
     4741 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     4801 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     4861 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgac ctcgaggggg
     4921 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     4981 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5041 gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5101 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5521 ctgccgctta ccggatacct gtccgccttt ctcttcggga agcgtggcgc tttctcatag
     5581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5641 cgaacccccc gttcagcccg accctgcgcc ttatccggta actatcgtct tgagtccaac
     5701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5821 agaacagtat ttggtatctg cgctctgctg aagccagtta cttcggaaaa agagttggta
     5881 gctcttgatc cggcaaacaa accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5941 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6001 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6061 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6121 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6361 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6421 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6661 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6781 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6841 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6961 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7021 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7081 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccactaaat tgtaagcgtt
     7141 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     7201 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     7261 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     7321 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     7381 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     7441 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     7501 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     7561 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     7621 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     7681 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     7741 gagcgcgcgt aatacgactc actatagggc gaattggagc tccaccgcgg tctttcggac
     7801 ttttgaaagt gatggtggtg ggggaaggat tcgaaccttc gaagtcgatg acggcagatt
     7861 tagagtctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7921 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
