LOCUS       sequence-315012-        6561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of CD44 proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6561)
  AUTHORS   Mielgo A, van Driel M, Bloem A, Landmann L, Gunthert U
  TITLE     A novel antiapoptotic mechanism based on interference of Fas 
            signaling by CD44 variant isoforms.
  JOURNAL   Cell Death Differ. 2006 Mar;13(3):465-77. doi: 
            10.1038/sj.cdd.4401763.
  PUBMED    16167069
REFERENCE   2  (bases 1 to 6561)
  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: "10.1038/sj.cdd"; 
            journalName: "Cell Death Differ"; date: "2006-03"; volume: "13"; 
            issue: "3"; pages: "465-77"
COMMENT     Sequence Label: pPGK-T7/2-CD44v2-10 (fl)
FEATURES             Location/Qualifiers
     source          1..6561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..500
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(36..59)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     433..452
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     intron          550..1122
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1130..1149
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1177..1196
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1177..1195
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3458..3467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3464..3466,3467..3469,3470..4183)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3464..3466,3467..3469,3470..4180)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3770..3789)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4117..4138
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4217..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4254..4273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4308..4327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4380..4484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4485..4553,4554..5345)
                     /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     complement(4703..4722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5516..6104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6005..6024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6258..6275
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6429..6448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
       61 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
      121 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
      181 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
      241 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
      301 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
      361 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
      421 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
      481 cctcatctcc gggcctttcg acccctaggc ttttgcaaaa agctccggat cgatcctgag
      541 aacttcaggg tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc
      601 atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg
      661 tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca
      721 ttgtctcctc ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat
      781 ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa
      841 tcactttttt ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga
      901 acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt
      961 ggaaatattc ttattggtag aaacaactac atcctggtca tcatcctgcc tttctcttta
     1021 tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc
     1081 ccctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     1141 gttattgtgc tgtctcatca ttttggcaaa gaattgtaat acgactcact atagggcgaa
     1201 ttccgctccg gacaccatgg acaagttttg gtggcacgca gcctggggac tctgcctcgt
     1261 gccgctgagc ctggcgcaga tcgatttgaa tataacctgc cgctttgcag gtgtattcca
     1321 cgtggagaaa aatggtcgct acagcatctc tcggacggag gccgctgacc tctgcaaggc
     1381 tttcaatagc accttgccca caatggccca gatggagaaa gctctgagca tcggatttga
     1441 gacctgcagg tatgggttca tagaagggca cgtggtgatt ccccggatcc accccaactc
     1501 catctgtgca gcaaacaaca caggggtgta catcctcaca tccaacacct cccagtatga
     1561 cacatattgc ttcaatgctt cagctccacc tgaagaagat tgtacatcag tcacagacct
     1621 gcccaatgcc tttgatggac caattaccat aactattgtt aaccgtgatg gcacccgcta
     1681 tgtccagaaa ggagaataca gaacgaatcc tgaagacatc taccccagca accctactga
     1741 tgatgacgtg agcagcggct cctccagtga aaggagcagc acttcaggag gttacatctt
     1801 ttacaccttt tctactgtac accccatccc agacgaagac agtccctgga tcaccgacag
     1861 cacagacaga atccctgcta ccactttgat gagcactagt gctacagcaa ctgagacagc
     1921 aaccaagagg caagaaacct gggattggtt ttcatggttg tttctaccat cagagtcaaa
     1981 gaatcatctt cacacaacaa cacaaatggc tggtacgtct tcaaatacca tctcagcagg
     2041 ctgggagcca aatgaagaaa atgaagatga aagagacaga cacctcagtt tttctggatc
     2101 aggcattgat gatgatgaag attttatctc cagcaccatt tcaaccacac cacgggcttt
     2161 tgaccacaca aaacagaacc aggactggac ccagtggaac ccaagccatt caaatccgga
     2221 ggtgctactt cagacaacca caaggatgac tgatgtagac agaaatggca ccactgctta
     2281 tgaaggaaac tggaacccag aagcacaccc tcccctcatt caccatgagc atcatgagga
     2341 agaagagacc ccacattcta caagcacaat ccaggcaact cctagtagta caacggaaga
     2401 aacagctacc cagaaggaac agtggtttgg caacagatgg catgagggat atcgccaaac
     2461 acccagagaa gactcccatt cgacaacagg gacagctgca gcctcagctc ataccagcca
     2521 tccaatgcaa ggaaggacag caccaagccc tgaggacagt tcctggactg atttcttcaa
     2581 cccaatctca caccccatgg gacgaggtca tcaagcagga agaaggatgg atatggactc
     2641 cagtcatagt acaacgcttc agcctactgc aaatccaaac acaggtttgg tggaagattt
     2701 ggacaggaca ggacctcttt caatgacaac gcagcagtgt aattctcaga gcttctctac
     2761 atcacatgaa ggcttggaag aagataaaga ccatccaaca acttctactc tgacatcaag
     2821 caataggaat gatgtcacag gtggaagaag agacccaaat cattctgaag gctcaactac
     2881 tttactggaa ggttatacct ctcattaccc acacacgaag gaaagcagga ccttcatccc
     2941 agtgacctca gctaagactg ggtcctttgg agttactgca gttactgttg gagattccaa
     3001 ctctaatgtc aatcgttcct tatcaggaga ccaagacaca ttccacccca gtggggggtc
     3061 ccataccact catggatctg aatcagatgg acactcacat gggagtcaag aaggtggagc
     3121 aaacacaacc tctggtccta taaggacacc ccaaattcca gaatggctga tcatcttggc
     3181 atccctcttg gccttggctt tgattcttgc agtttgcatt gcagtcaaca gtcgaagaag
     3241 gtgtgggcag aagaaaaagc tagtgatcaa cagtggcaat ggagctgtgg aggacagaaa
     3301 gccaagtgga ctcaacggag aggccagcaa gtctcaggaa atggtgcatt tggcgaacaa
     3361 ggagtcgtca gaaactccag accagtttat gacagctgat gagacaagga acctgcagaa
     3421 tgtggacatg aagattgggg tagatccccg gccggtcgcc accatggtga gcaagggcga
     3481 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3541 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     3601 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3661 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3721 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3781 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3841 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3901 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3961 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4021 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4081 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     4141 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgagatcca
     4201 gatcttatta aagcagaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4261 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4321 actcatcaat gtatcttatc atgtctggtc gagcaggtgg cacttttcgg ggaaatgtgc
     4381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4441 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4501 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4561 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4621 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4681 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4741 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4801 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4861 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4921 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4981 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5041 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5101 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5161 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5221 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5281 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5341 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5401 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5641 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5881 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5941 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6001 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6061 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6121 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6181 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6241 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6301 aaccgcctct ccccgcgcgt tggccgattc atttcgactt ctgaggcgga aagaaccagg
     6361 ccaatatggg atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     6421 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6481 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6541 ccctgaatga acaattctac c
//
