LOCUS       sequence-276417-        6075 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 6075)
  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 6075)
  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-CD44v6-10 (fl)
FEATURES             Location/Qualifiers
     source          1..6075
                     /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      2972..2981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2978..2980,2981..2983,2984..3697)
                     /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(2978..2980,2981..2983,2984..3694)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3284..3303)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3631..3652
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3731..3852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3768..3787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3822..3841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3894..3998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3999..4067,4068..4859)
                     /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(4217..4236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5030..5618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5519..5538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5772..5789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5943..5962)
                     /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 aaccgcgatg 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 ccatccaggc aactcctagt agtacaacgg aagaaacagc
     1921 tacccagaag gaacagtggt ttggcaacag atggcatgag ggatatcgcc aaacacccag
     1981 agaagactcc cattcgacaa cagggacagc tgcagcctca gctcatacca gccatccaat
     2041 gcaaggaagg acaacaccaa gcccagagga cagttcctgg actgatttct tcaacccaat
     2101 ctcacacccc atgggacgag gtcatcaagc aggaagaagg atggatatgg actccagtca
     2161 tagtacaacg cttcagccta ctgcaaatcc aaacacaggt ttggtggaag atttggacag
     2221 gacaggacct ctttcaatga caacgcagca gagtaattct cagagcttct ctacatcaca
     2281 tgaaggcttg gaagaagata aagaccatcc aacaacttct actctgacat caagcaatag
     2341 gaatgatgtc acaggtggaa gaagagaccc aaatcattct gaaggctcaa ctactttact
     2401 ggaaggttat acctctcatt acccacacac gaaggaaagc aggaccttca tcccagtgac
     2461 ctcagctaag actgggtcct ttggagttac tgcagttact gttggagatt ccaactctaa
     2521 tgtcaatcgt tccttatcag gagaccaaga cacattccac cccagtgggg ggtcccatac
     2581 cactcatgga tctgaatcag atggacactc acatgggagt caagaaggtg gagcaaacac
     2641 aacctctggt cctataagga caccccaaat tccagaatgg ctgatcatct tggcatccct
     2701 cttggccttg gctttgattc ttgcagtttg cattgcagtc aacagtcgaa gaaggtgtgg
     2761 gcagaagaaa aagctagtga tcaacagtgg caatggagct gtggaggaca gaaagccaag
     2821 tggactcaac ggagaggcca gcaagtctca ggaaatggtg catttggtga acaaggagtc
     2881 gtcagaaact ccagaccagt ttatgacagc tgatgagaca aggaacctgc agaatgtgga
     2941 catgaagatt ggggtagatc cccggccggt cgccaccatg gtgagcaagg gcgaggagct
     3001 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3061 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     3121 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     3181 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     3241 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3301 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     3361 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     3421 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     3481 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3541 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     3601 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3661 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgaga tccagatctt
     3721 attaaagcag aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3841 caatgtatct tatcatgtct ggtcgagcag gtggcacttt tcggggaaat gtgcgcggaa
     3901 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3961 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4021 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4081 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4141 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4201 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4261 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4321 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4381 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4441 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4501 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4561 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4621 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4681 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4741 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4801 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4861 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4921 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4981 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5041 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5101 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5161 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5221 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5281 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5341 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5401 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5461 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5521 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5581 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5641 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5701 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5761 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5821 ctctccccgc gcgttggccg attcatttcg acttctgagg cggaaagaac caggccaata
     5881 tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5941 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     6001 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6061 atgaacaatt ctacc
//
