LOCUS       Exported                7851 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral vector for expression of eGFP-P2A-Puro casette from a CMV
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pLCE-P2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7851)
  AUTHORS   Patil A, Manzano M, Gottwein E
  TITLE     CK1alpha and IRF4 are essential and independent effectors of 
            immunomodulatory drugs in primary effusion lymphoma.
  JOURNAL   Blood. 2018 Aug 9;132(6):577-586. doi: 10.1182/blood-2018-01-828418.
            Epub 2018 Jun 28.
  PUBMED    29954751
REFERENCE   2  (bases 1 to 7851)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7851
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    37..70
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    126..714
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(179..199)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(597..608)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(717..733)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(718..734)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1243..1423
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(1485..2073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1565..1584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2244..3104)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2867..2886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3105..3209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3284..3303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3475..3854
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3856..4054
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             4072..4252
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4299..4424
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4921..5154
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5339..5383
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5681..5798
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        5920..6223
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6224..6427
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      6466..6475
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6472..7188
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             6472..7188
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6517..6538)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6778..6797)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7125..7146
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7198..7254
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7255..7851
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7748..7768
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 aatatgcggc cgccagagaa ttcgtcgagg gacctaataa cttcgtatag catacattat
       61 acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta
      121 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
      181 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
      241 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
      301 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
      361 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
      421 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
      481 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
      541 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
      601 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
      661 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
      721 accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
      781 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
      841 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
      901 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
      961 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     1021 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     1081 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     1141 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     1201 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     1261 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     1321 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     1381 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc
     1441 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1501 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1561 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1621 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1681 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1741 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1801 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1861 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1921 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1981 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2041 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2101 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2161 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2221 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2281 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2341 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2401 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2461 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2521 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2581 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2641 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2701 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2761 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2821 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2881 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2941 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3001 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3061 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3121 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3181 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3241 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     3301 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     3361 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     3421 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     3481 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     3541 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     3601 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3661 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     3721 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     3781 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     3841 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     3901 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     3961 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4021 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
     4081 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4141 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4201 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     4261 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     4321 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     4381 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     4441 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     4501 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     4561 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     4621 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     4681 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     4741 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     4801 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     4861 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     4921 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     4981 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     5041 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     5101 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     5161 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     5221 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     5281 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     5341 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     5401 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     5461 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     5521 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     5581 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     5641 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     5701 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     5761 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     5821 gagatccagt ttggttagta ccgggcccga gaggatcctt agttattaat agtaatcaat
     5881 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5941 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6001 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6061 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6121 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6181 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     6241 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6301 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6361 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6421 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc
     6481 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     6541 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     6601 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     6661 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     6721 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     6781 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     6841 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     6901 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     6961 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     7021 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     7081 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     7141 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg atccggcgca
     7201 acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgaccgag
     7261 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     7321 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     7381 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     7441 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     7501 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     7561 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     7621 ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     7681 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     7741 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     7801 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg a
//
