LOCUS       sequence-101451-        5991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a SV40
            promoter and LacZ module. Compatible with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  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 5991)
  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 SV40 LacZ L5-L2
FEATURES             Location/Qualifiers
     source          1..5991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(11..110)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(140..159)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(141..159)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(164..180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(164..180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(282..301)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             293..1102
                     /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(360..379)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1249..1837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1738..1757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1991..2008
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2167..2194)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2286..2372)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2435..2452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2436..2452
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2469..2564
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2569..2589)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2574..2903
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2754..2889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2924..5968
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
ORIGIN
        1 ctagattact acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
       61 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
      121 gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
      181 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
      241 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
      301 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
      361 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
      421 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
      481 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
      541 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
      601 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
      661 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
      721 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
      781 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
      841 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
      901 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
      961 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     1021 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     1081 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     1141 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     1201 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1261 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1321 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1381 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1501 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1561 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1621 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1681 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1801 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1861 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1921 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     1981 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     2041 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     2101 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     2161 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     2221 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     2281 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     2341 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     2401 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
     2461 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     2521 tgagcaatgc ttttttataa tgccaacttt gtatacaaaa gttgctgtgg aatgtgtgtc
     2581 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2641 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2701 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2761 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2821 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2881 ttggaggcct aggcttttgc aaaaagcttc catgatagat cccgtcgttt tacaacgtcg
     2941 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     3001 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3061 gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct
     3121 ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg
     3181 ttacgatgcg cccatctaca ccaacgtaac ctatcccatt acggtcaatc cgccgtttgt
     3241 tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct
     3301 acaggaaggc cagacgcgaa ttatttttga tggcgttaac tcggcgtttc atctgtggtg
     3361 caacgggcgc tgggtcggtt acggccagga cagtcgtttg ccgtctgaat ttgacctgag
     3421 cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgtt ggagtgacgg
     3481 cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg acgtctcgtt
     3541 gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct ttaatgatga
     3601 tttcagccgc gctgtactgg aggctgaagt tcagatgtgc ggcgagttgc gtgactacct
     3661 acgggtaaca gtttctttat ggcagggtga aacgcaggtc gccagcggca ccgcgccttt
     3721 cggcggtgaa attatcgatg agcgtggtgg ttatgccgat cgcgtcacac tacgtctgaa
     3781 cgtcgaaaac ccgaaactgt ggagcgccga aatcccgaat ctctatcgtg cggtggttga
     3841 actgcacacc gccgacggca cgctgattga agcagaagcc tgcgatgtcg gtttccgcga
     3901 ggtgcggatt gaaaatggtc tgctgctgct gaacggcaag ccgttgctga ttcgaggcgt
     3961 taaccgtcac gagcatcatc ctctgcatgg tcaggtcatg gatgagcaga cgatggtgca
     4021 ggatatcctg ctgatgaagc agaacaactt taacgccgtg cgctgttcgc attatccgaa
     4081 ccatccgctg tggtacacgc tgtgcgaccg ctacggcctg tatgtggtgg atgaagccaa
     4141 tattgaaacc cacggcatgg tgccaatgaa tcgtctgacc gatgatccgc gctggctacc
     4201 ggcgatgagc gaacgcgtaa cgcgaatggt gcagcgcgat cgtaatcacc cgagtgtgat
     4261 catctggtcg ctggggaatg aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg
     4321 gatcaaatct gtcgatcctt cccgcccggt gcagtatgaa ggcggcggag ccgacaccac
     4381 ggccaccgat attatttgcc cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc
     4441 tgtgccgaaa tggtccatca aaaaatggct ttcgctacct ggagagacgc gcccgctgat
     4501 cctttgcgaa tacgcccacg cgatgggtaa cagtcttggc ggtttcgcta aatactggca
     4561 ggcgtttcgt cagtatcccc gtttacaggg cggcttcgtc tgggactggg tggatcagtc
     4621 gctgattaaa tatgatgaaa acggcaaccc gtggtcggct tacggcggtg attttggcga
     4681 tacgccgaac gatcgccagt tctgtatgaa cggtctggtc tttgccgacc gcacgccgca
     4741 tccagcgctg acggaagcaa aacaccagca gcagtttttc cagttccgtt tatccgggca
     4801 aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc gataacgagc tcctgcactg
     4861 gatggtggcg ctggatggta agccgctggc aagcggtgaa gtgcctctgg atgtcgctcc
     4921 acaaggtaaa cagttgattg aactgcctga actaccgcag ccggagagcg ccgggcaact
     4981 ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca tggtcagaag ccgggcacat
     5041 cagcgcctgg cagcagtggc gtctggcgga aaacctcagt gtgacgctcc ccgccgcgtc
     5101 ccacgccatc ccgcatctga ccaccagcga aatggatttt tgcatcgagc tgggtaataa
     5161 gcgttggcaa tttaaccgcc agtcaggctt tctttcacag atgtggattg gcgataaaaa
     5221 acaactgctg acgccgctgc gcgatcagtt cacccgtgca ccgctggata acgacattgg
     5281 cgtaagtgaa gcgacccgca ttgaccctaa cgcctgggtc gaacgctgga aggcggcggg
     5341 ccattaccag gccgaagcag cgttgttgca gtgcacggca gatacacttg ctgatgcggt
     5401 gctgattacg accgctcacg cgtggcagca tcaggggaaa accttattta tcagccggaa
     5461 aacctaccgg attgatggta gtggtcaaat ggcgattacc gttgatgttg aagtggcgag
     5521 cgatacaccg catccggcgc ggattggcct gaactgccag ctggcgcagg tagcagagcg
     5581 ggtaaactgg ctcggattag ggccgcaaga aaactatccc gaccgcctta ctgccgcctg
     5641 ttttgaccgc tgggatctgc cattgtcaga catgtatacc ccgtacgtct tcccgagcga
     5701 aaacggtctg cgctgcggga cgcgcgaatt gaattatggc ccacaccagt ggcgcggcga
     5761 cttccagttc aacatcagcc gctacagtca acagcaactg atggaaacca gccatcgcca
     5821 tctgctgcac gcggaagaag gcacatggct gaatatcgac ggtttccata tggggattgg
     5881 tggcgacgac tcctggagcc cgtcagtatc ggcggaattc cagctgagcg ccggtcgcta
     5941 ccattaccag ttggtctggt gtcaaaaagc ggccgctcga gtctaaaagg t
//
