LOCUS       sequence-197604-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human CD95L leucine zipper in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  AUTHORS   Rad S M AH, Poudel A, Tan GMY, McLellan AD
  TITLE     Promoter choice: Who should drive the CAR in T cells?
  JOURNAL   PLoS One. 2020 Jul 24;15(7):e0232915. doi: 
            10.1371/journal.pone.0232915. eCollection 2020.
  PUBMED    32706785
REFERENCE   2  (bases 1 to 6117)
  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"; journalName: "PLoS One.";
            date: "2020-07-24"; volume: "15(7):e0232915. doi"; pages: " 
            10.1371/journal.pone.0232915. eCollection 2020"
COMMENT     Sequence Label: pcDNA3.1(-)-LZ-CD95L
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..105
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(757..774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    763..987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1033..1461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1120..1139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1330..1351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1470..1490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1475..1804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1655..1790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1717..1736
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1871..2665
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1925..1944)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2535..2554
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2839..2960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2876..2895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2930..2949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3009..3025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3009..3025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3022..3044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3033..3049
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(join(3057..3063,3064..3081,3082..3087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3102..3123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3240..3257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3411..3996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3491..3510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4167..4958,4959..5027))
                     /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     4790..4809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5028..5132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5207..5226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5398..5777
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5778..5981
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5932..5952
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6026..6045
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6026..6044
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aattcggcct ctgaggccac catgcgaagg atgcaacttc ttctgctcat cgcactttcc
       61 ttggcgctgg tgacaaatag tcatcaccac catcaccatc atcacggaga tcggatgaaa
      121 cagatcgaag acaagattga agagattctg tcaaaaatat atcatatcga aaatgagata
      181 gcgcggatca agaagttgat cggcgaaaga acttctgggg ggtctggagg aactggcgga
      241 agcgggggaa caggtggaag cccccctgaa aaaaaggagc tgaggaaagt ggcccattta
      301 acaggcaagt ccaactcaag gtccatgcct ctggaatggg aagacaccta tggaattgtc
      361 ctgctttctg gagtgaagta taagaagggt ggccttgtga tcaatgaaac tgggctgtac
      421 tttgtatatt ccaaagtata cttccggggt caatcttgca acaacctgcc cctgagccac
      481 aaggtctaca tgaggaactc taagtatccc caggatctgg tgatgatgga ggggaagatg
      541 atgagctact gcactactgg gcagatgtgg gcccgcagca gctacctggg ggcagtgttc
      601 aatcttacca gtgctgatca tttatatgtc aacgtatctg agctctctct ggtcaatttt
      661 gaggaatctc agacgttttt cggcttatat aagctctaag gcctgtcagg ccggatccga
      721 gctcggtacc aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc
      781 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
      841 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
      901 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
      961 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     1021 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1081 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1141 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1201 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1261 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1321 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     1381 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1441 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     1501 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     1561 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1621 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     1681 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     1741 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     1801 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     1861 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     1921 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     1981 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     2041 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     2101 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     2161 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     2221 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     2281 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     2341 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     2401 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     2461 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     2521 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     2581 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     2641 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     2701 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     2761 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     2821 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2881 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2941 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     3001 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3061 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3121 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3181 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     3961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5161 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     5221 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     5281 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     5341 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     5401 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     5461 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5521 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5581 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5641 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5701 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5761 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5821 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5881 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5941 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
     6001 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
     6061 agcgtttaaa cgggccctct agactcgagc ggccgccact gtgctggata tctgcag
//
