LOCUS       sequence-418347-        5003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for in vitro transcription template preparation, insert T7 promoter,
            TP53 translation initiation site P0 to +670 and F-luc from 
            pGL3-Basic, into pUC19. P0~670 is confusing, BLAST is recommended..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5003)
  TITLE     RBM24 interacts with RPL3 and regulates RPL3-eIF6 interaction
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5003)
  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            "Unpublished"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: pUC-T7-hTP53-P0~670-FLuc
FEATURES             Location/Qualifiers
     source          1..5003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             690..2342
                     /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(756..774)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2386..2402)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2386..2402)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2399..2421)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2410..2426
                     /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(2434..2440,2441..2458,2459..2464))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2479..2500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2617..2634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2788..3376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2868..2887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3547..4338,4339..4407))
                     /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     4170..4189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4408..4512)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4580..4598
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4636..4658)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4758..4777
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4971..4993
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4985..5002
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4986..5002
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattctaata cgactcacta tagtcatggc gactgtccag ctttgtgcca ggagcctcgc
       61 aggggttgat gggattgggg ttttcccctc ccatgtgctc aagactggcg ctaaaagttt
      121 tgagcttctc aaaagtctag agccaccgtc cagggagcag gtagctgctg ggctccgggg
      181 acactttgcg ttcgggctgg gagcgtgctt tccacgacgg tgacacgctt ccctggattg
      241 gcagccagac tgccttccgg gtcactgcca tggaggagcc gcagtcagat cctagcgtcg
      301 agccccctct gagtcaggaa acattttcag acctatggaa actacttcct gaaaacaacg
      361 ttctgtcccc cttgccgtcc caagcaatgg atgatttgat gctgtccccg gacgatattg
      421 aacaatggtt cactgaagac ccaggtccag atgaagctcc cagaatgcca gaggctgctc
      481 cccccgtggc ccctgcacca gcagctccta caccggcggc ccctgcacca gccccctcct
      541 ggcccctgtc atcttctgtc ccttcccaga aaacctacca gggcagctac ggtttccgtc
      601 tgggcttctt gcattctggg acagccaagt ctgtgacttg cacgtactcc cctgccctca
      661 acaagatgtt ttgccaactg gccaagacca tggaagacgc caaaaacata aagaaaggcc
      721 cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga
      781 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca
      841 tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
      901 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
      961 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
     1021 aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa
     1081 aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta
     1141 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
     1201 atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga
     1261 caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc
     1321 ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa
     1381 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta
     1441 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
     1501 agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc
     1561 tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     1621 aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt
     1681 tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca gctattctga
     1741 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     1801 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt
     1861 gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
     1921 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
     1981 acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc
     2041 ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag
     2101 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     2161 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     2221 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     2281 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt
     2341 aattctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     2401 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2461 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2521 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4561 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4621 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4681 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4741 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     4801 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     4861 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     4921 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     4981 acgttgtaaa acgacggcca gtg
//
