LOCUS       sequence-176932-        6829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  AUTHORS   Godar S, Ince TA, Bell GW, Feldser D, Donaher JL, Bergh J, Liu A, 
            Miu K, Watnick RS, Reinhardt F, McAllister SS, Jacks T, Weinberg RA
  TITLE     Growth-inhibitory and tumor- suppressive functions of p53 depend on 
            its repression of CD44 expression.
  JOURNAL   Cell. 2008 Jul 11. 134(1):62-73.
  PUBMED    18614011
REFERENCE   2  (bases 1 to 6829)
  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: "Cell. 2008 
            Jul 11. 134(1):62-73."
COMMENT     Sequence Label: CD44P pGL3
FEATURES             Location/Qualifiers
     source          1..6829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    4..125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(15..34)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     69..88
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(373..390)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(544..1132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(624..643)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1303..2094,2095..2163))
                     /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     1926..1945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2164..2268)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2295..2750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2382..2401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2592..2613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2881..2929
                     /note="synthetic polyadenylation signal"
     misc_feature    2943..3034
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2983..3002
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      5134..5143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5140..6792
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(5206..5224)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
       61 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      121 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      181 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg
      241 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
      301 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
      361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      961 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1261 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1321 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1381 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1441 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1501 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1561 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1621 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1681 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1741 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1801 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1861 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1921 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1981 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2041 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2101 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2161 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2221 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2281 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2341 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2401 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2461 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2521 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2581 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2641 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2701 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc
     2761 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     2821 gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc
     2881 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
     2941 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
     3001 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcttacgcg
     3061 tgctagcctt aatccatgct gttcgtcact acactgtact gcctgtggat gacttacttg
     3121 tccctgtagt ttcatctgaa gaattcctcc tccttttcct ttgaggcctg cctcaaatat
     3181 cacttcccct gtgaagactg cctggtgttc tccaggagag agtgtgactc ccttctctag
     3241 gaatggtagc accccaaaca cacacatttt gcagcatatt tcaccttgca tggtaatggc
     3301 ctgcttgtga gttttattcc gtaccagagg gtgagggctc tgaagatagc gccaggtctt
     3361 atttacctcg ataccccaca acactcatta catgtctgat gaatgaatgc atagggggat
     3421 ggctgggtgc atttctctca acttttcaat ttcttgaaat aaataacaaa atcttacctt
     3481 ccctcagaag tcctggcatg gttcctttca tcttgccaca gccactgata atcactttca
     3541 ttttctgtgt aactcaccag gcaagaagtc catgcagatt tacttttagt agttcacatg
     3601 acaaataaat actgcgtttg atttccaaac attaaaccat agtatattat agatagatat
     3661 agagttatca ttcaaagtat gatatttcaa tctcaaaagg cttcccctga agaatattac
     3721 aaactcttcc tctctttaag atctgctggg taggaaagat gggagaaaat gaattaatgt
     3781 ttacacagaa aggaggataa tgggggcaaa aataatagat gaacgtatgg gtggatgaga
     3841 gaatggataa aatgataggt ggatatgttg atcttggaca gatgggaaat gagtggatat
     3901 atcaataaac agatatgtgg gtggatgggt ggagaagagg atggtggatg gttgtggttt
     3961 tatgaagaga tgtgaaaaag gaagtgtgga atgatggatg agaagttgta tgggaagatg
     4021 aatagaagaa taggtggttg aataaattaa aaggtgtgtg gttggatgaa tgaatgagtg
     4081 ggatgataga tggacctaag tggttagtgg atggacagga ggatggatgg atgtgagagc
     4141 cccagaagga cataaggaaa gatgggtgga tagatggatg ggcggatgga aggatattta
     4201 ggaggatgaa tgagcatgtg tgtggagaga ggtgcccatt cacactggct tgaacacatg
     4261 ggttagctga gccaaatgcc agccctatga caggccatca gtagctttcc ctgagctgtt
     4321 ctgccaagaa gctaaaattc attcaagcca tgtggacttg ttattgaggg gaaaaagaat
     4381 gagctctccc tctttccact tggaagattc accaactccc cacccctcac tccccactgt
     4441 gggcacggag gcactgcgcc acccagggca agacctcgcc ctctctccag ctcctctccc
     4501 aggatatcca acatcctgtg aaacccagag atcttgctcc agccggattc agagaaattt
     4561 agcgggaaag gagaggccaa aggctgaacc caatggtgca aggttttacg gttcggtcat
     4621 cctctgtcct gacgccgcgg ggccagcggg agaagaaagc cagtgcgtct ctgggcgcag
     4681 gggccagtgg ggctcggagg cacaggcacc ccgcgacact ccaggttccc cgacccacgt
     4741 ccctggcagc cccgattatt tacagcctca gcagagcacg gggcgggggc agaggggccc
     4801 gcccgggagg gctgctactt cttaaaacct ctgcgggctg cttagtcaca gccccccttg
     4861 cttgggtgtg tccttcgctc gctccctccc tccgtcttag gtcactgttt tcaacctcga
     4921 ataaaaactg cagccaactt ccgaggcagc ctcattgccc agcggacccc agcctctgcc
     4981 aggttcggtc cgccatcctc gtcccgtcct ccgccggccc ctgccccgcg cccagggatc
     5041 ctccagctcc tttcgcccgc gccctccgtt cgctccggac accctcgaga tctgcgatct
     5101 aagtaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
     5161 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
     5221 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
     5281 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
     5341 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
     5401 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
     5461 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
     5521 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
     5581 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
     5641 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
     5701 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
     5761 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
     5821 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     5881 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     5941 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     6001 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     6061 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     6121 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     6181 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     6241 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     6301 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     6361 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     6421 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     6481 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     6541 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     6601 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     6661 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     6721 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     6781 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagaca
//
