LOCUS       sequence-473393-        5347 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express HiBiT-TP53(Y220C) in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5347)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5347)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HiBiT-TP53(Y220C) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1134..2318
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPCEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSDV"
     primer_bind     complement(2426..2444)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2481..2602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2492..2511)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2546..2565
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2847..3204
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2847..3038
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2870..2890)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3055..3190
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3219..3266
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3298..4092
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3352..3371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3962..3981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4156..4204
                     /note="synthetic polyadenylation signal"
     rep_origin      4361..4906
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4807..4826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5023..5040
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatggagg
     1141 agccgcagtc agatcctagc gtcgagcccc ctctgagtca ggaaacattt tcagacctat
     1201 ggaaactact tcctgaaaac aacgttctgt cccccttgcc gtcccaagca atggatgatt
     1261 tgatgctgtc cccggacgat attgaacaat ggttcactga agacccaggt ccagatgaag
     1321 ctcccagaat gccagaggct gctccccccg tggcccctgc accagcagct cctacaccgg
     1381 cggcccctgc accagccccc tcctggcccc tgtcatcttc tgtcccttcc cagaaaacct
     1441 accagggcag ctacggtttc cgtctgggct tcttgcattc tgggacagcc aagtctgtga
     1501 cttgcacgta ctcccctgcc ctcaacaaga tgttttgcca actggccaag acctgccctg
     1561 tgcagctgtg ggttgattcc acacccccgc ccggcacccg cgtccgcgcc atggccatct
     1621 acaagcagtc acagcacatg acggaggttg tgaggcgctg cccccaccat gagcgctgct
     1681 cagatagcga tggtctggcc cctcctcagc atcttatccg agtggaagga aatttgcgtg
     1741 tggagtattt ggatgacaga aacacttttc gacatagtgt ggtggtgccc tgtgagccgc
     1801 ctgaggttgg ctctgactgt accaccatcc actacaacta catgtgtaac agttcctgca
     1861 tgggcggcat gaaccggagg cccatcctca ccatcatcac actggaagac tccagtggta
     1921 atctactggg acggaacagc tttgaggtgc gtgtttgtgc ctgtcctggg agagaccggc
     1981 gcacagagga agagaatctc cgcaagaaag gggagcctca ccacgagctg cccccaggga
     2041 gcactaagcg agcactgccc aacaacacca gctcctctcc ccagccaaag aagaaaccac
     2101 tggatggaga atatttcacc cttcagatcc gtgggcgtga gcgcttcgag atgttccgag
     2161 agctgaatga ggccttggaa ctcaaggatg cccaggctgg gaaggagcca ggggggagca
     2221 gggctcactc cagccacctg aagtccaaaa agggtcagtc tacctcccgc cataaaaaac
     2281 tcatgttcaa gacagaaggg cctgactcag acgtttaaac gaattcgggc tcggtacccg
     2341 gggatcctct agagtcgacc tgcaggcatg caagctgatc cggctgctaa caaagcccga
     2401 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcg
     2461 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2521 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2581 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2641 gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     2701 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     2761 gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct
     2821 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     2881 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2941 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3001 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3061 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3121 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3181 tttggaggcc taggcttttg caaaaagctt aattaactgt tgacaattaa tcatcggcat
     3241 agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag gcagatcatg
     3301 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3361 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3421 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3481 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3541 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3601 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3661 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3721 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3781 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3841 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3901 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3961 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4021 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4081 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4141 catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     4201 tgtgaatcga tagcgataag gatccgctca tgcgatgata agctgtcaaa catgagaatt
     4261 acaacttata tcgtatgggg ctgacttcag gtgctacatt tgaagagata aattgcactg
     4321 aaatctagaa atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc
     4381 gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc
     4441 tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac
     4501 ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa
     4561 ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga
     4621 tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc
     4681 ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca
     4741 taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc
     4801 gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga
     4861 gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc
     4921 ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt
     4981 tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag
     5041 gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc
     5101 ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac
     5161 actccgctag cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcatgaga
     5221 ttacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     5281 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     5341 gagatct
//
