LOCUS       V000780                 9377 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000780
VERSION     V000780
KEYWORDS    pMuLE Lenti Dest iRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9377)
  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 9377)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9377)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Clin
            Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             94..327
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    399..533
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      539..674
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(707..725)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(735..751)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      896..1351
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1380..1484
                     /label="AmpR promoter"
     CDS             1485..2342
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2516..3104
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     3258..3275
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    3392..3413
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3428..3458
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3466..3482
                     /label="lac operator"
                     /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)."
     primer_bind     3490..3506
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3527..3545
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3573..3800
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3801..3981
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4025..4150
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    4643..4876
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5060..5104
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             5253..5294
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    5365..5482
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    5537..5661
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5686..5716
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             5770..6426
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             6771..7073
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(7117..7241)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    7272..7860
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        7874..8373
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             8417..9364
                     /label="iRFP713"
                     /note="phytochrome-based near-infrared fluorescent protein,
                     originally termed iRFP (Filonov et al., 2011)"
ORIGIN
        1 cgcgtatgca tctcgagggc ccggtacctt taagaccaat gacttacaag gcagctgtag
       61 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctagctcac tcccaacgaa
      121 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
      181 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      241 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
      301 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
      361 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
      421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
      481 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
      541 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
      601 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
      661 cttttttgga ggcctaggct tttgcgtcga gacgtaccca attcgcccta tagtgagtcg
      721 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      781 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      841 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcgacgcg
      901 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
      961 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1021 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1081 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1141 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1201 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1261 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1321 aattttaaca aaatattaac gtttacaatt tcccaggtgg cacttttcgg ggaaatgtgc
     1381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1441 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1501 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1561 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1621 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1681 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1741 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1801 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1861 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1921 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1981 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2041 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2101 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2161 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2221 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2281 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2341 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2401 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2641 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     2701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2881 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2941 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3001 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3061 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3121 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3181 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3241 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3301 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3361 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     3421 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     3481 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     3541 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     3601 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     3661 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     3721 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     3781 cctagctcga tacaataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     3841 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     3901 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     3961 cagtgtggaa aatctctagc agtggcgccc gaacagggac ctgaaagcga aagggaaacc
     4021 agagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     4081 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     4141 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     4201 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     4261 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     4321 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     4381 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     4441 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     4501 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     4561 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     4621 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     4681 gcactatggg cgcagcctca atgacgctga cggtacaggc cagacaatta ttgtctggta
     4741 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     4801 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     4861 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     4921 tgccttggaa tgctagttgg agtaataaat ctctggaaca gattggaatc acacgacctg
     4981 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     5041 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     5101 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     5161 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     5221 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     5281 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     5341 agtgaacgga tctcgacggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc
     5401 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     5461 aattacaaaa attcaaaatt ttattccagt gtggtggaat tgagtattcc agtgtggtgg
     5521 aattctgcag atatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
     5581 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     5641 acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct
     5701 tccggctcgt ataatgtgtg gattttgagt taggatccgg cgagattttc aggagctaag
     5761 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     5821 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     5881 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     5941 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     6001 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     6061 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     6121 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     6181 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     6241 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     6301 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat
     6361 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     6421 ggggcgtaaa gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
     6481 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
     6541 ggtgtgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
     6601 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
     6661 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
     6721 gtttattgaa atgaacggct cttttgctga cgagaacagg gactggtgaa atgcagttta
     6781 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     6841 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     6901 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     6961 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     7021 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     7081 caggctccgt tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt
     7141 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     7201 tcattttacg tttctcgttc agctttcttg tacaaagtgg ttgatatcca gcacagtggc
     7261 ggccgctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     7321 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     7381 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     7441 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     7501 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     7561 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     7621 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     7681 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     7741 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     7801 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     7861 ctggaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
     7921 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca
     7981 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact
     8041 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa
     8101 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag
     8161 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
     8221 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
     8281 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
     8341 ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt accacactcc
     8401 tgcatctgcc gccaccatgg cggaaggatc cgtcgccagg cagcctgacc tcttgacctg
     8461 cgacgatgag ccgatccata tccccggtgc catccaaccg catggactgc tgctcgccct
     8521 cgccgccgac atgacgatcg ttgccggcag cgacaacctt cccgaactca ccggactggc
     8581 gatcggcgcc ctgatcggcc gctctgcggc cgatgtcttc gactcggaga cgcacaaccg
     8641 tctgacgatc gccttggccg agcccggggc ggccgtcgga gcaccgatca ctgtcggctt
     8701 cacgatgcga aaggacgcag gcttcatcgg ctcctggcat cgccatgatc agctcatctt
     8761 cctcgagctc gagcctcccc agcgggacgt cgccgagccg caggcgttct tccgccgcac
     8821 caacagcgcc atccgccgcc tgcaggccgc cgaaaccttg gaaagcgcct gcgccgccgc
     8881 ggcgcaagag gtgcggaaga ttaccggctt cgatcgggtg atgatctatc gcttcgcctc
     8941 cgacttcagc ggcgaagtga tcgcagagga tcggtgcgcc gaggtcgagt caaaactagg
     9001 cctgcactat cctgcctcaa ccgtgccggc gcaggcccgt cggctctata ccatcaaccc
     9061 ggtacggatc attcccgata tcaattatcg gccggtgccg gtcaccccag acctcaatcc
     9121 ggtcaccggg cggccgattg atcttagctt cgccatcctg cgcagcgtct cgcccgtcca
     9181 tctggaattc atgcgcaaca taggcatgca cggcacgatg tcgatctcga ttttgcgcgg
     9241 cgagcgactg tggggattga tcgtttgcca tcaccgaacg ccgtactacg tcgatctcga
     9301 tggccgccaa gcctgcgagc tagtcgccca ggttctggcc tggcagatcg gcgtgatgga
     9361 agagtgagtc gacgcga
//
