LOCUS       sequence-101456-        5361 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 5361)
  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 5361)
  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 L5-L2
FEATURES             Location/Qualifiers
     source          1..5361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(54..153)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(184..202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(207..223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(207..223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(325..344)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             336..1145
                     /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(403..422)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1292..1880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1781..1800
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2034..2051
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2210..2237)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2329..2415)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2464..2486
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2478..2495
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2479..2495
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2512..2607
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        2635..3014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3015..3218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3169..3189
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3263..3281
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3356..4384
                     /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             4397..5332
                     /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             4445..5338
                     /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 ggtacccagc
       61 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
      121 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagggc ggccgcgata
      181 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
      241 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
      301 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
      361 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
      421 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
      481 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
      541 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
      601 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
      661 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
      721 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
      781 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
      841 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
      901 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
      961 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     1021 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     1081 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     1141 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     1201 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     1261 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1321 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1381 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1441 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1501 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1561 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1621 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1681 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1741 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1801 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1861 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1921 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1981 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2041 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2101 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2161 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     2221 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     2281 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     2341 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     2401 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
     2461 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga
     2521 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta
     2581 taatgccaac tttgtataca aaagttggcg atgtacgggc cagatatacg cgttgacatt
     2641 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2701 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2761 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2821 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2881 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2941 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     3001 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     3061 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     3121 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     3181 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     3241 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
     3301 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgagccg ccaccatgtc
     3361 caatttactg accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga
     3421 ggttcgcaag aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg
     3481 gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa
     3541 atggtttccc gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg
     3601 tctggcagta aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc
     3661 cgggctgcca cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa
     3721 agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt
     3781 cgaccaggtt cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc
     3841 atttctgggg attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt
     3901 taaagatatc tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac
     3961 gctggttagc accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga
     4021 gcgatggatt tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt
     4081 cagaaaaaat ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga
     4141 agggattttt gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag
     4201 atacctggcc tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc
     4261 tggagtttca ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat
     4321 gaactatatc cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg
     4381 cgatctcgag ccatctgctg gagacatgag agctgccaac ctttggccaa gcccgctcat
     4441 gatcaaacgc tctaagaaga acagcctggc cttgtccctg acggccgacc agatggtcag
     4501 tgccttgttg gatgctgagc cccccatact ctattccgag tatgatccta ccagaccctt
     4561 cagtgaagct tcgatgatgg gcttactgac caacctggca gacagggagc tggttcacat
     4621 gatcaactgg gcgaagaggg tgccaggctt tgtggatttg accctccatg atcaggtcca
     4681 ccttctagaa tgtgcctggc tagagatcct gatgattggt ctcgtctggc gctccatgga
     4741 gcacccagtg aagctactgt ttgctcctaa cttgctcttg gacaggaacc agggaaaatg
     4801 tgtagagggc atggtggaga tcttcgacat gctgctggct acatcatctc ggttccgcat
     4861 gatgaatctg cagggagagg agtttgtgtg cctcaaatct attattttgc ttaattctgg
     4921 agtgtacaca tttctgtcca gcaccctgaa gtctctggaa gagaaggacc atatccaccg
     4981 agtcctggac aagatcacag acactttgat ccacctgatg gccaaggcag gcctgaccct
     5041 gcagcagcag caccagcggc tggcccagct cctcctcatc ctctcccaca tcaggcacat
     5101 gagtaacaaa ggcatggagc atctgtacag catgaagtgc aagaacgtgg tgcccctcta
     5161 tgacctgctg ctggaggcgg cggacgccca ccgcctacat gcgcccacta gccgtggagg
     5221 ggcatccgtg gaggagacgg accaaagcca cttggccact gcgggctcta cttcatcgca
     5281 ttccttgcaa aagtattaca tcacggggga ggcagagggt ttccctgcca cagcttgatg
     5341 aagatctgag ctccctggcg g
//
