LOCUS       sequence-101457-        5471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a CMV 
            promoter and CreERT2 module. Compatible with MultiSite Gateway 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5471)
  AUTHORS   Albers J, Danzer C, Rechsteiner M, Lehmann H, Brandt LP, Hejhal T, 
            Catalano A, Busenhart P, Goncalves AF, Brandt S, Bode PK, 
            Bode-Lesniewska B, Wild PJ, Frew IJ
  TITLE     A versatile modular vector system for rapid combinatorial mammalian 
            genetics.
  JOURNAL   J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743.
  PUBMED    25751063
REFERENCE   2  (bases 1 to 5471)
  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: "J Clin 
            Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743."
COMMENT     Sequence Label: pMuLE ENTR CMV CreERT2 R4-R3
FEATURES             Location/Qualifiers
     source          1..5471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    54..177
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(233..251)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(256..272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(256..272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(374..393)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             385..1194
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(452..471)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1341..1929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1830..1849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2083..2100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2259..2286)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2378..2464)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2513..2535
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2527..2544
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2528..2544
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2594..2718)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        2745..3124
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3125..3328
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3279..3299
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3373..3391
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3466..4494
                     /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="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             4507..5442
                     /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             4555..5448
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
ORIGIN
        1 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga ggacaacttt
       61 gtataataaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
      121 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatgaat
      181 caactactta gatggtatta gtgacctgta ctgcagggcg gccgcgatat cccctatagt
      241 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
      301 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
      361 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
      421 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
      481 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
      541 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
      601 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
      661 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
      721 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
      781 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
      841 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
      901 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
      961 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     1021 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     1081 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     1141 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     1201 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     1261 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     1321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1921 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1981 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2041 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2101 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2161 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2221 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     2281 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     2341 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     2401 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     2461 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     2521 cgacgttgta aaacgacggc cagtcttaag ctcgggcccc tacaggtcac taataccatc
     2581 taagtagttg attcatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt
     2641 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac
     2701 ttttctatac aaagttggcg atgtacgggc cagatatacg cgttgacatt gattattgac
     2761 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     2821 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     2881 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     2941 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3001 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3061 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3121 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     3181 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     3241 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     3301 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
     3361 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat
     3421 ccactagtaa cggccgccag tgtgctggaa ttctgagccg ccaccatgtc caatttactg
     3481 accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga ggttcgcaag
     3541 aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg gaaaatgctt
     3601 ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa atggtttccc
     3661 gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg tctggcagta
     3721 aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc cgggctgcca
     3781 cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa agaaaacgtt
     3841 gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt cgaccaggtt
     3901 cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc atttctgggg
     3961 attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt taaagatatc
     4021 tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac gctggttagc
     4081 accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga gcgatggatt
     4141 tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt cagaaaaaat
     4201 ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga agggattttt
     4261 gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag atacctggcc
     4321 tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc tggagtttca
     4381 ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat gaactatatc
     4441 cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg cgatctcgag
     4501 ccatctgctg gagacatgag agctgccaac ctttggccaa gcccgctcat gatcaaacgc
     4561 tctaagaaga acagcctggc cttgtccctg acggccgacc agatggtcag tgccttgttg
     4621 gatgctgagc cccccatact ctattccgag tatgatccta ccagaccctt cagtgaagct
     4681 tcgatgatgg gcttactgac caacctggca gacagggagc tggttcacat gatcaactgg
     4741 gcgaagaggg tgccaggctt tgtggatttg accctccatg atcaggtcca ccttctagaa
     4801 tgtgcctggc tagagatcct gatgattggt ctcgtctggc gctccatgga gcacccagtg
     4861 aagctactgt ttgctcctaa cttgctcttg gacaggaacc agggaaaatg tgtagagggc
     4921 atggtggaga tcttcgacat gctgctggct acatcatctc ggttccgcat gatgaatctg
     4981 cagggagagg agtttgtgtg cctcaaatct attattttgc ttaattctgg agtgtacaca
     5041 tttctgtcca gcaccctgaa gtctctggaa gagaaggacc atatccaccg agtcctggac
     5101 aagatcacag acactttgat ccacctgatg gccaaggcag gcctgaccct gcagcagcag
     5161 caccagcggc tggcccagct cctcctcatc ctctcccaca tcaggcacat gagtaacaaa
     5221 ggcatggagc atctgtacag catgaagtgc aagaacgtgg tgcccctcta tgacctgctg
     5281 ctggaggcgg cggacgccca ccgcctacat gcgcccacta gccgtggagg ggcatccgtg
     5341 gaggagacgg accaaagcca cttggccact gcgggctcta cttcatcgca ttccttgcaa
     5401 aagtattaca tcacggggga ggcagagggt ttccctgcca cagcttgatg aagatctgag
     5461 ctccctggcg g
//
