LOCUS       sequence-353218-        5595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression/secretion in mammalian cells. CD44 splice variant
            10 (exons 2-5, 11, 15-16), A20-E649..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5595)
  AUTHORS   Gileadi, et al
  TITLE     TREAT-AD plasmids 2021
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5595)
  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: 
            "unpublished"
COMMENT     Sequence Label: CD44U-c010
FEATURES             Location/Qualifiers
     source          1..5595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2820..2837
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2949..2968)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3014..3069
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3068..3087)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3430..3446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3430..3446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3443..3465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3454..3470
                     /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(3478..3484,3485..3502,3503..3508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3523..3544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3673..3690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3844..4432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3924..3943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4603..5394,5395..5463))
                     /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     5226..5245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5464..5568)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs grggcggaga ggkgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga attcaagctt gccaccatgg ggatccttcc cagccctggg atgcctgcgc
     1801 tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct gaaaccggtg
     1861 cgcagatcga tttgaatata acctgccgct ttgcaggtgt attccacgtg gagaaaaatg
     1921 gtcgctacag catctctcgg acggaggccg ctgacctctg caaggctttc aatagcacct
     1981 tgcccacaat ggcccagatg gagaaagctc tgagcatcgg atttgagacc tgcaggtatg
     2041 ggttcataga agggcatgtg gtgattcccc ggatccaccc caactccatc tgtgcagcaa
     2101 acaacacagg ggtgtacatc ctcacatcca acacctccca gtatgacaca tattgcttca
     2161 atgcttcagc tccacctgaa gaagattgta catcagtcac agacctgccc aatgcctttg
     2221 atggaccaat taccataact attgttaacc gtgatggcac ccgctatgtc cagaaaggag
     2281 aatacagaac gaatcctgaa gacatctacc ccagcaaccc tactgatgat gacgtgagca
     2341 gcggctcctc cagtgaaagg agcagcactt caggaggtta catcttttac accttttcta
     2401 ctgtacaccc catcccagac gaagacagtc cctggatcac cgacagcaca gacagaatcc
     2461 ctgctaccaa taggaatgat gtcacaggtg gaagaagaga cccaaatcat tctgaaggct
     2521 caactacttt actggaaggt tatacctctc attacccaca cacgaaggaa agcaggacct
     2581 tcatcccagt gacctcagct aagactgggt cctttggagt tactgcagtt actgttggag
     2641 attccaactc taatgtcaat cgttccttat caggagacca agacacattc caccccagtg
     2701 gggggtccca taccactcat ggatctgaat cagatggaca ctcacatggg agtcaagaag
     2761 gtggagcaaa cacaacctct ggtcctataa ggacacccca aattccagaa ggtaccaagc
     2821 accaccatca ccatcactaa tgatcactcg agactagtat cgcgataatt cactcctcag
     2881 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     2941 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     3001 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     3061 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     3121 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     3181 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     3241 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     3301 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     3361 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     3421 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     3601 cgatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5581 aaaagtgcca cctgg
//
