LOCUS       sequence-471893-        5809 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express LgBiT-p53 Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5809)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5809)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: LgBiT-p53 Control Vector
FEATURES             Location/Qualifiers
     source          1..5809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     228..248
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    516..637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(553..572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     607..626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     674..692
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(800..1984)
                     /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
                     SPDDIEQWFTEDPGPDEAPRMPEAAPRVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSDV"
     primer_bind     complement(2528..2547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2529..2547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2589..2608)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2591..2723
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2859..3062)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2888..2908)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3063..3442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3887..3904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4021..4566)
                     /direction=LEFT
                     /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     complement(4101..4120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4723..4771
                     /note="synthetic polyadenylation signal"
     CDS             complement(4835..5629)
                     /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(4946..4965)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5556..5575
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5661..5708)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(5723..5809,1..271))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5737..5809,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      241 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
      301 ggttatttca ggccatggtg ctgcgcagat ccgcgtatgc ggtgtgaaat accgcacaga
      361 tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg
      421 attttaccac atttgtagag gttttacttg cttaaaaaaa cctcccacat ctccccctga
      481 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
      541 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
      601 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg aagcggccgc
      661 cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg
      721 ggctttgtta gcagccggat cagcttgcat gcctgcaggt cgactctaga ggatccccgg
      781 gtaccgagcc cgaattcgtt taaacgtctg agtcaggccc ttctgtcttg aacatgagtt
      841 ttttatggcg ggaggtagac tgaccctttt tggacttcag gtggctggag tgagccctgc
      901 tcccccctgg ctccttccca gcctgggcat ccttgagttc caaggcctca ttcagctctc
      961 ggaacatctc gaagcgctca cgcccacgga tctgaagggt gaaatattct ccatccagtg
     1021 gtttcttctt tggctgggga gaggagctgg tgttgttggg cagtgctcgc ttagtgctcc
     1081 ctgggggcag ctcgtggtga ggctcccctt tcttgcggag attctcttcc tctgtgcgcc
     1141 ggtctctccc aggacaggca caaacacgca cctcaaagct gttccgtccc agtagattac
     1201 cactggagtc ttccagtgtg atgatggtga ggatgggcct ccggttcatg ccgcccatgc
     1261 aggaactgtt acacatgtag ttgtagtgga tggtggtaca gtcagagcca acctcaggcg
     1321 gctcataggg caccaccaca ctatgtcgaa aagtgtttct gtcatccaaa tactccacac
     1381 gcaaatttcc ttccactcgg ataagatgct gaggaggggc cagaccatcg ctatctgagc
     1441 agcgctcatg gtgggggcag cgcctcacaa cctccgtcat gtgctgtgac tgcttgtaga
     1501 tggccatggc gcggacgcgg gtgccgggcg ggggtgtgga atcaacccac agctgcacag
     1561 ggcaggtctt ggccagttgg caaaacatct tgttgagggc aggggagtac gtgcaagtca
     1621 cagacttggc tgtcccagaa tgcaagaagc ccagacggaa accgtagctg ccctggtagg
     1681 ttttctggga agggacagaa gatgacaggg gccaggaggg ggctggtgca ggggccgccg
     1741 gtgtaggagc tgctggtgca ggggccacgc ggggagcagc ctctggcatt ctgggagctt
     1801 catctggacc tgggtcttca gtgaaccatt gttcaatatc gtccggggac agcatcaaat
     1861 catccattgc ttgggacggc aagggggaca gaacgttgtt ttcaggaagt agtttccata
     1921 ggtctgaaaa tgtttcctga ctcagagggg gctcgacgct aggatctgac tgcggctcct
     1981 ccatggcgat cgcaccgctc gagcctccac ctccgctccc gccaccaccg gagctcccac
     2041 tgttgatggt tactcggaac agcatggagc cgtcgggggt gatcaggcgc tcgtcgataa
     2101 ttttgttgcc gttccacagg gtccctgtta cagtgatctt tttgccgtcg aacacggcga
     2161 tgccttcata cggccgtccg aaatagttca gcatgttcgg cgtaaccccg tcgattacca
     2221 gtgtgccata gggcaggatc accttaaagt gatgatcatc cacagggtac accaccttaa
     2281 acacctcttc gatctgggcc atttggtcgg cgctcagacc ttcatacggg atgatgacat
     2341 ggatgtcgat cttcagggca ttttcaccgc tccggacaat cctttggatc ggagttacgg
     2401 acacggcgag attctgcagc aaactggaca cacctccctg ttcaaggact tggtccaggt
     2461 tgtaggcggc tgtctgttcc cagtccccaa cgaaatcttc gagtgtgaag accatggtga
     2521 gcgctagccc tatagtgagt cgtattaata ctctagcctt aagagctgta attgaactgg
     2581 gagtggacac ctgtggagag aaaggcaaag tggatgtcag taagaccaat aggtgcctat
     2641 cagaaacgca agagtcttct ctgtctcgac aagcccagtt tctattggtc tccttaaacc
     2701 tgtcttgtaa ccttgatact tacctgccca gtgcctcacg accaacttct gcagcttaag
     2761 ttcgagactg ttgtgtcaga agcactgact gcgttagcaa tttaactgtg ataaactacc
     2821 gcaataaagc ttctagtgat ctgacggttc actaaaccag ctctgcttat atagacctcc
     2881 caccgtacac gcctaccgcc catttgcgtc aacggggcgg ggttattacg acattttgga
     2941 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     3001 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattgg tgtactgcca aaaccgcatc
     3061 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccgtaag
     3121 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     3181 ggacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     3241 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     3301 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     3361 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     3421 taactagtca ataatcaatg ccaacatggc ggtcatattg gacatgagcc aatataaatg
     3481 tacatattat gatatagata caacgtatgc aatggccaat agccaatatt gatttatgct
     3541 atataaccaa tgaataatat ggctaatggc caatattgaa gatctcgatc ctctacgccg
     3601 gacgcatcgt ggccggagta ctgcggttta tctcgatggc tacgagggca gacagtaagt
     3661 ggatttacca taatccctta attgtacgca ccgctaaaac gcgtaatctc atgacggggt
     3721 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     3781 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     3841 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     3901 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     3961 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     4021 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     4081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     4141 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     4201 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     4261 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4321 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     4381 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     4441 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     4501 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     4561 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     4621 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     4681 acagcttatc atcgcatgag cggatcctta tcgctatcga ttcacacaaa aaaccaacac
     4741 acagatgtaa tgaaaataaa gatattttat tgcggccatc gtgatggcag gttgggcgtc
     4801 gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     4861 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     4921 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     4981 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     5041 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     5101 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     5161 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     5221 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     5281 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     5341 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     5401 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     5461 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     5521 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     5581 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg atctgcctcc
     5641 tgggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc cgatgattaa
     5701 ttgtcaacag ttaattaagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     5761 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaat
//
