LOCUS       sequence-471839-        5457 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express TP53-HiBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5457)
  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: TP53-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5457
                     /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             1065..2243
                     /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
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(2400..2418)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2455..2576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2466..2485)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2520..2539
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2821..3178
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2821..3012
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2844..2864)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3029..3164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3193..3240
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3272..4066
                     /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(3326..3345)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3936..3955
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4130..4178
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4417..5005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4497..4516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5121..5404
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 ggctagcgat cgccatggag gagccgcagt
     1081 cagatcctag cgtcgagccc cctctgagtc aggaaacatt ttcagaccta tggaaactac
     1141 ttcctgaaaa caacgttctg tcccccttgc cgtcccaagc aatggatgat ttgatgctgt
     1201 ccccggacga tattgaacaa tggttcactg aagacccagg tccagatgaa gctcccagaa
     1261 tgccagaggc tgctcccccc gtggcccctg caccagcagc tcctacaccg gcggcccctg
     1321 caccagcccc ctcctggccc ctgtcatctt ctgtcccttc ccagaaaacc taccagggca
     1381 gctacggttt ccgtctgggc ttcttgcatt ctgggacagc caagtctgtg acttgcacgt
     1441 actcccctgc cctcaacaag atgttttgcc aactggccaa gacctgccct gtgcagctgt
     1501 gggttgattc cacacccccg cccggcaccc gcgtccgcgc catggccatc tacaagcagt
     1561 cacagcacat gacggaggtt gtgaggcgct gcccccacca tgagcgctgc tcagatagcg
     1621 atggtctggc ccctcctcag catcttatcc gagtggaagg aaatttgcgt gtggagtatt
     1681 tggatgacag aaacactttt cgacatagtg tggtggtgcc ctatgagccg cctgaggttg
     1741 gctctgactg taccaccatc cactacaact acatgtgtaa cagttcctgc atgggcggca
     1801 tgaaccggag gcccatcctc accatcatca cactggaaga ctccagtggt aatctactgg
     1861 gacggaacag ctttgaggtg cgtgtttgtg cctgtcctgg gagagaccgg cgcacagagg
     1921 aagagaatct ccgcaagaaa ggggagcctc accacgagct gcccccaggg agcactaagc
     1981 gagcactgcc caacaacacc agctcctctc cccagccaaa gaagaaacca ctggatggag
     2041 aatatttcac ccttcagatc cgtgggcgtg agcgcttcga gatgttccga gagctgaatg
     2101 aggccttgga actcaaggat gcccaggctg ggaaggagcc aggggggagc agggctcact
     2161 ccagccacct gaagtccaaa aagggtcagt ctacctcccg ccataaaaaa ctcatgttca
     2221 agacagaagg gcctgactca gacgtttctc aaggcagttc aggtggctcg agcggtgtga
     2281 gcggctggcg gctgttcaag aagattagct aaggccgcga ctctagagtc gacctgcagg
     2341 catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     2401 cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat
     2461 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2521 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2581 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa
     2641 gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca
     2701 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg
     2761 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
     2821 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2881 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2941 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3001 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3061 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3121 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3181 gcttaattaa ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     3241 aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg
     3301 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3361 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3421 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3481 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3541 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3601 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3661 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3721 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3781 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3841 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3901 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3961 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4021 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4081 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc
     4141 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatcct
     4201 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     4261 gcaccgtttc tgcggactgg ctttctaccc ggtatcagct cactcaaagg cggtaatacg
     4321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4981 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5101 cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg
     5161 gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt
     5221 accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt
     5281 tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt
     5341 tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca
     5401 tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
