LOCUS       sequence-190929-        6642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutant p53 encoded by AS-30D rat hepatoma cell line.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6642)
  AUTHORS   Mathupala SP, Heese C, Pedersen PL
  TITLE     Glucose catabolism in cancer cells. The type II hexokinase promoter 
            contains functionally active response elements for the tumor 
            suppressor p53.
  JOURNAL   J Biol Chem. 1997 Sep 5;272(36):22776-80.
  PUBMED    9278438
REFERENCE   2  (bases 1 to 6642)
  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: "J Biol 
            Chem."; date: "1997-09-5"; volume: "272(36)"; pages: "22776-80"
COMMENT     Sequence Label: pCI-Neo-p53mut
FEATURES             Location/Qualifiers
     source          1..6642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        211..590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        591..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     799..823
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          963..1095
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1078..1097
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1140..1158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1175..1184
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2419..2540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2430..2449)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2484..2503
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2726..3181
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2813..2832)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3023..3044
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3298..3655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3298..3489
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3321..3341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3506..3641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3706..4500
                     /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(3760..3779)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4370..4389
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4564..4612
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4653..4672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4772..4794
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4832..4850)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4918..5022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5023..5091,5092..5883)
                     /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     complement(5241..5260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6054..6642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6543..6562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 gctcgacaga tcttcaatat tggccattag ccatattatt cattggttat atagcataaa
      121 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
      181 ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt tattaatagt
      241 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      301 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      361 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      421 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
      481 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
      541 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      601 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      661 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      721 gtcgtaacaa ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa gctttattgc
      841 ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa
      901 cttaagctgc agtgactctc ttaaggtagc cttgcagaag ttggtcgtga ggcactgggc
      961 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
     1021 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
     1081 cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt
     1141 aatacgactc actataggct agcctcgaga attcgccacc atggaggatt cacagtcgga
     1201 tatgagcatc gagctccctc tgagtcagga gacattttca tgcttatgga aacttcttcc
     1261 tccagatgat attctgccca ccacagcgac agggtcacct aattccatgg aagatctgtt
     1321 cctgccccag gatgttgcag agttgttaga aggcccagag gaagccctcc aagtgtcagc
     1381 tcctgcagca caggaacctg gaactgaggc ccctgcaccc gtggcccctg cttcagctac
     1441 accgtggcct ctgtcatctt ccgtcccttc tcaaaaaact taccaaagca actatggctt
     1501 ccacctgggc ttcctgcagt cagggacagc caagtctgtt atgtgcacgt actcaatttc
     1561 cctcaataag ctgttctgcc agctggcgaa gacatgccct gtgcagttgt gggtcacctc
     1621 cacacctcca cctggtaccc gtgtccgtgc catggccatc tacaagaagt cacaacacat
     1681 gactgaggtc gtgagacgct gcccccacca tgagcgttgc tctgatggtg acggcctggc
     1741 tcctccccaa catcttatcc gggtggaagg aaatccgtat gctgagtatc tggacgacag
     1801 gcagactttt cggcacagcg tggtggtacc gtatgagcca cctgaggtcg gctccgacta
     1861 taccactatc cactacaagt acatgtgcaa cagctcctgc atggggggca tgaaccgccg
     1921 gcccatcctt accatcatca cgctgggaga ctccagtggg aatcttctgg gacgggacag
     1981 ctttgaggtt cgtgtttgtg cctgtcctgg gagagaccgt cggacagagg aagaaaattt
     2041 ccgcaaaaaa gaagagcatt gcccggagct gcccccaggg agtgcaaaga gagcactgcc
     2101 caccagcaca agctcctctc cccagcaaaa ggaaaaacca ctcgatggag aatatttcac
     2161 ccttaagatc cgtgggcgtg agcgcttcga gatgttccga gagctgaatg aggccttgga
     2221 attaaaggat gcccgtgctg ccgaggagtc aggagacagc agggctcact ccagctaccc
     2281 gaagaccaag aagggccagt ctacgtcccg ccataaaaaa ccaatgatca agaaagtggg
     2341 gcctgactca gactgatcta gagtcgaccc gggcggccgc ttccctttag tgagggttaa
     2401 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2461 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2521 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2581 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     2641 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2701 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2761 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2821 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2881 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2941 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3001 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3061 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3121 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3181 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     3241 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3301 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3361 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3421 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3481 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3541 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3601 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3661 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     3721 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3781 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3841 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3901 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3961 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4021 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4081 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4141 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4201 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4261 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4321 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4381 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4441 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4501 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     4561 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     4621 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4681 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4741 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4801 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4861 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5101 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5161 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5221 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5281 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5341 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5401 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5461 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5521 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5581 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5641 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5701 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5761 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5821 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5881 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5941 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aa
//
