LOCUS       Exported                4365 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a CMV 
            promoter and iRFP module. Compatible with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pMuLE ENTR CMV iRFP R4-R3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4365)
  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 4365)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4365
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    9..132
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(188..206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(211..227)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(211..227)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(329..348)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             340..1149
                     /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(407..426)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1296..1884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1785..1804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2038..2055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2214..2241
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2333..2419
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2468..2490
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2482..2499
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2483..2499
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2549..2673)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        2700..3079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3080..3283
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3234..3254
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3328..3346
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3381..3390
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3387..4337
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
ORIGIN
        1 ctagaggaca actttgtata ataaagttga acgagaaacg taaaatgata taaatatcaa
       61 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
      121 ccagtcacta tgaatcaact acttagatgg tattagtgac ctgtactgca gggcggccgc
      181 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
      241 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
      301 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
      361 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
      421 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
      481 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
      541 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
      601 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
      661 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
      721 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
      781 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
      841 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
      901 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
      961 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     1021 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     1081 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     1141 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     1201 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     1261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1441 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2041 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     2101 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     2161 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     2221 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     2281 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     2341 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     2401 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     2461 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gcccctacag
     2521 gtcactaata ccatctaagt agttgattca tagtgactgg atatgttgtg ttttacagta
     2581 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta
     2641 cgtttctcgt tcaacttttc tatacaaagt tggcgatgta cgggccagat atacgcgttg
     2701 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     2761 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     2821 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2881 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2941 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     3001 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     3061 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     3121 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     3181 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     3241 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
     3301 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg
     3361 gtaccgagct cggatctgcc gccaccatgg cggaaggatc cgtcgccagg cagcctgacc
     3421 tcttgacctg cgacgatgag ccgatccata tccccggtgc catccaaccg catggactgc
     3481 tgctcgccct cgccgccgac atgacgatcg ttgccggcag cgacaacctt cccgaactca
     3541 ccggactggc gatcggcgcc ctgatcggcc gctctgcggc cgatgtcttc gactcggaga
     3601 cgcacaaccg tctgacgatc gccttggccg agcccggggc ggccgtcgga gcaccgatca
     3661 ctgtcggctt cacgatgcga aaggacgcag gcttcatcgg ctcctggcat cgccatgatc
     3721 agctcatctt cctcgagctc gagcctcccc agcgggacgt cgccgagccg caggcgttct
     3781 tccgccgcac caacagcgcc atccgccgcc tgcaggccgc cgaaaccttg gaaagcgcct
     3841 gcgccgccgc ggcgcaagag gtgcggaaga ttaccggctt cgatcgggtg atgatctatc
     3901 gcttcgcctc cgacttcagc ggcgaagtga tcgcagagga tcggtgcgcc gaggtcgagt
     3961 caaaactagg cctgcactat cctgcctcaa ccgtgccggc gcaggcccgt cggctctata
     4021 ccatcaaccc ggtacggatc attcccgata tcaattatcg gccggtgccg gtcaccccag
     4081 acctcaatcc ggtcaccggg cggccgattg atcttagctt cgccatcctg cgcagcgtct
     4141 cgcccgtcca tctggaattc atgcgcaaca taggcatgca cggcacgatg tcgatctcga
     4201 ttttgcgcgg cgagcgactg tggggattga tcgtttgcca tcaccgaacg ccgtactacg
     4261 tcgatctcga tggccgccaa gcctgcgagc tagtcgccca ggttctggcc tggcagatcg
     4321 gcgtgatgga agagtgagtc gacgcggccg ctcgagccta agctt
//
