LOCUS       sequence-239598-        7857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of non-targeting control shRNA #4 
            from a miR30 embedded casette in the 3'UTR of ZsGreen-P2A-Puro 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7857)
  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 7857)
  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/blood-2018-01-828418"; journalName: "Blood"; date: 
            "2018-08-9- 9"; volume: "132"; issue: "6"; pages: "577-586"
COMMENT     Sequence Label: pZIP-hCMV-ZsGreen-P2A-Puro-NT4
FEATURES             Location/Qualifiers
     source          1..7857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             411..668
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    715..840
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1337..1570
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1755..1799
                     /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    2062..2177
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        2235..2538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2539..2742
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2693..2713
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2739..2763
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2833..3525
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     CDS             3535..3591
                     /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             3592..4188
                     /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     4085..4105
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     ncRNA           4209..4330
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4358..4372
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4400..4526
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    4560..5148
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4613..4633)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5031..5042)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5355..5588
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5679..5699)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5684..6013
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5864..5999
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5926..5945
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     rep_origin      complement(6102..6690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6182..6201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6861..7652,7653..7721))
                     /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     7484..7503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7722..7826)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcctcg cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accagctagc gaggcgtggc
      421 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc
      481 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
      541 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
      601 tctgttgtgt gactctggta actagagatc cctccgaccc ttttagtcag tgtggaaaat
      661 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
      721 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
      781 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
      841 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
      901 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
      961 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     1021 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     1081 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     1141 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     1201 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1261 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1321 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1381 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1441 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1501 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1561 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1621 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1681 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1741 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     1801 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1861 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     1921 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     1981 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2041 acggatctcg acggtatcgc ctttaaaaga aaagggggga ttggggggta cagtgcaggg
     2101 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2161 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttatcgataa
     2221 gcttgggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2281 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2341 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2401 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     2461 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     2521 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     2581 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2641 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2701 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     2761 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg actctagcta
     2821 gagtaccggt ccatggccca gtccaagcac ggcctgacca aggagatgac catgaagtac
     2881 cgcatggagg gctgcgtgga cggccacaag ttcgtgatca ccggcgaggg catcggctac
     2941 cccttcaagg gcaagcaggc catcaacctg tgcgtggtgg agggcggccc cttgcccttc
     3001 gccgaggaca tcttgtccgc cgccttcatg tacggcaacc gcgtgttcac cgagtacccc
     3061 caggacatcg tcgactactt caagaactcc tgccccgccg gctacacctg ggaccgctcc
     3121 ttcctgttcg aggacggcgc cgtgtgcatc tgcaacgccg acatcaccgt gagcgtggag
     3181 gagaactgca tgtaccacga gtccaagttc tacggcgtga acttccccgc cgacggcccc
     3241 gtgatgaaga agatgaccga caactgggag ccctcctgcg agaagatcat ccccgtgccc
     3301 aagcagggca tcttgaaggg cgacgtgagc atgtacctgc tgctgaagga cggtggccgc
     3361 ttgcgctgcc agttcgacac cgtgtacaag gccaagtccg tgccccgcaa gatgcccgac
     3421 tggcacttca tccagcacaa gctgacccgc gaggaccgca gcgacgccaa gaaccagaag
     3481 tggcacctga ccgagcacgc catcgcctcc ggctccgcct tgcccggatc cggcgcaaca
     3541 aacttctctc tgctgaaaca agccggagat gtcgaagaga atcctggacc gaccgagtac
     3601 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     3661 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     3721 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     3781 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     3841 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     3901 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     3961 ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     4021 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     4081 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     4141 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagc ggccgcaatg
     4201 gatcctgttt gaatgaggct tcagtacttt acagaatcgt tgcctgcaca tcttggaaac
     4261 acttgctggg attacttctt caggttaacc caacagaagg ctcgagaagg tatattgctg
     4321 ttgacagtga gcgaaggcag aagtatgcaa agcattagtg aagccacaga tgtaatgctt
     4381 tgcatacttc tgcctgtgcc tactgcctcg gacttcaagg ggctacttta ggagcaatta
     4441 tcttgtttac taaaactgaa taccttgcta tctctttgat acatttttac aaagctgaat
     4501 taaaatggta taaattaaat cactttagct tgcagatcta cgcgtggcgc gcctggaaca
     4561 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4621 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4681 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4741 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4801 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4861 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4921 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     4981 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5041 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5101 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt
     5161 ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     5221 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     5281 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5341 agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     5401 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5461 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5521 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5581 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     5641 aatatcagag agtgagaggc cttgacattg tttaaactgg aatgtgtgtc agttagggtg
     5701 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5761 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5821 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5881 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5941 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6001 aggcttttgc aaaaagctcc cgggagcttg tatatccatt acatgtgagc aaaaggccag
     6061 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6421 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6481 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6541 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6601 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6661 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6721 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6781 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6841 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6901 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6961 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7021 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7081 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7141 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7201 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7261 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7321 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7381 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7441 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7501 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7561 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7621 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7681 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7741 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7801 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
