LOCUS       sequence-242486-        7830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of ZsGreen-P2A-Puro casette from a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7830)
  AUTHORS   Patil A, Manzano M, Gottwein E
  TITLE     Genome-wide CRISPR screens reveal genetic mediators of cereblon 
            modulator toxicity in primary effusion lymphoma.
  JOURNAL   Blood Adv. 2019 Jul 23;3(14):2105-2117. doi: 
            10.1182/bloodadvances.2019031732.
  PUBMED    31300418
REFERENCE   2  (bases 1 to 7830)
  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"; doi: 
            "10.1182/bloodadvances"; journalName: "Blood Adv"; date: 
            "2019-07-23- 23"; volume: "3"; issue: "14"; pages: "2105-2117"
COMMENT     Sequence Label: pLC-ZsGreen-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..7830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..66
                     /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             67..663
                     /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     560..580
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(700..733)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    789..1377
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(842..862)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1260..1271)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1380..1396)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1381..1397)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1906..2086
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2148..2736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2228..2247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2907..3698,3699..3767))
                     /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     3530..3549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3768..3872)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3947..3966)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4138..4517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4519..4717
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             4735..4915
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4962..5087
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5584..5817
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6002..6046
                     /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    6344..6461
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        6586..6889
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6890..7093
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      7132..7141
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7138..7830
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
ORIGIN
        1 ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct
       61 ggaccgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
      121 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
      181 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
      241 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
      301 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
      361 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
      421 gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg
      481 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
      541 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
      601 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
      661 tgaaatatgc ggccgccaga gaattcgtcg agggacctaa taacttcgta tagcatacat
      721 tatacgaagt tatacatgtt taagggttcc ggttccacta ggtacaattc gatatcaagc
      781 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
      841 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
      901 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
      961 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     1021 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     1081 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1141 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     1201 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     1261 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     1321 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     1381 gataccgtcg acctcgatcg agacctagaa aaacatggag caatcacaag tagcaataca
     1441 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     1501 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     1561 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     1621 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca
     1681 ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     1741 gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg
     1801 agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc
     1861 ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta
     1921 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1981 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2041 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat gtgagcaaaa
     2101 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2161 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2221 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2281 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2341 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2401 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2461 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2521 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2581 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2641 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2701 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2761 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2821 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2881 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2941 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3001 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3061 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3121 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3181 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3241 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3301 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3361 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3421 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3481 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3541 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3601 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3661 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3721 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3781 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3841 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3901 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     3961 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     4021 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     4081 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     4141 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     4201 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4261 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4321 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4381 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4441 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4501 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4561 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4621 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4681 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
     4741 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     4801 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     4861 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     4921 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     4981 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     5041 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     5101 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     5161 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     5221 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     5281 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     5341 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     5401 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     5461 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     5521 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     5581 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     5641 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     5701 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     5761 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     5821 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     5881 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     5941 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     6001 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     6061 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     6121 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     6181 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     6241 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     6301 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     6361 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     6421 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     6481 gcagagatcc agtttggtta gtaccgggcc cgagagttaa ttaattagtt attaatagta
     6541 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6601 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6661 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6721 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6781 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6841 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     6901 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6961 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     7021 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     7081 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
     7141 gcccagtcca agcacggcct gaccaaggag atgaccatga agtaccgcat ggagggctgc
     7201 gtggacggcc acaagttcgt gatcaccggc gagggcatcg gctacccctt caagggcaag
     7261 caggccatca acctgtgcgt ggtggagggc ggccccttgc ccttcgccga ggacatcttg
     7321 tccgccgcct tcatgtacgg caaccgcgtg ttcaccgagt acccccagga catcgtcgac
     7381 tacttcaaga actcctgccc cgccggctac acctgggacc gctccttcct gttcgaggac
     7441 ggcgccgtgt gcatctgcaa cgccgacatc accgtgagcg tggaggagaa ctgcatgtac
     7501 cacgagtcca agttctacgg cgtgaacttc cccgccgacg gccccgtgat gaagaagatg
     7561 accgacaact gggagccctc ctgcgagaag atcatccccg tgcccaagca gggcatcttg
     7621 aagggcgacg tgagcatgta cctgctgctg aaggacggtg gccgcttgcg ctgccagttc
     7681 gacaccgtgt acaaggccaa gtccgtgccc cgcaagatgc ccgactggca cttcatccag
     7741 cacaagctga cccgcgagga ccgcagcgac gccaagaacc agaagtggca cctgaccgag
     7801 cacgccatcg cctccggctc cgccttgccc
//
