LOCUS       Exported                6323 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses human p53 deltaN and mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pIRES2-mCherry-p53 deltaN
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6323)
  AUTHORS   Lin SC, Karoly ED, Taatjes DJ
  TITLE     The human DeltaNp53 isoform triggers metabolic and gene expression 
            changes that activate mTOR and alter mitochondrial function.
  JOURNAL   Aging Cell. 2013 Oct;12(5):863-72. doi: 10.1111/acel.12108. Epub 
            2013 Jun 25.
  PUBMED    23734707
REFERENCE   2  (bases 1 to 6323)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6323
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1703..2276
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1726..2278
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1870..1887)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2097..2116
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2278..2988
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2423..2441)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2691..2710
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3111..3232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3202..3221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3239..3694)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3376..3397)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3588..3607
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3721..3825
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3827..4184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3850..3870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4035..4170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4097..4116
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4219..5013
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4273..4292)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4883..4902
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5201..5220)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5245..5292
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5621..6209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6110..6129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgatat ggatgatttg atgctgtccc cggacgatat tgaacaatgg
      661 ttcactgaag acccaggtcc agatgaagct cccagaatgc cagaggctgc tccccccgtg
      721 gcccctgcac cagcagctcc tacaccggcg gcccctgcac cagccccctc ctggcccctg
      781 tcatcttctg tcccttccca gaaaacctac cagggcagct acggtttccg tctgggcttc
      841 ttgcattctg ggacagccaa gtctgtgact tgcacgtact cccctgccct caacaagatg
      901 ttttgccaac tggccaagac ctgccctgtg cagctgtggg ttgattccac acccccgccc
      961 ggcacccgcg tccgcgccat ggccatctac aagcagtcac agcacatgac ggaggttgtg
     1021 aggcgctgcc cccaccatga gcgctgctca gatagcgatg gtctggcccc tcctcagcat
     1081 cttatccgag tggaaggaaa tttgcgtgtg gagtatttgg atgacagaaa cacttttcga
     1141 catagtgtgg tggtgcccta tgagccgcct gaggttggct ctgactgtac caccatccac
     1201 tacaactaca tgtgtaacag ttcctgcatg ggcggcatga accggaggcc catcctcacc
     1261 atcatcacac tggaagactc cagtggtaat ctactgggac ggaacagctt tgaggtgcgt
     1321 gtttgtgcct gtcctgggag agaccggcgc acagaggaag agaatctccg caagaaaggg
     1381 gagcctcacc acgagctgcc cccagggagc actaagcgag cactgcccaa caacaccagc
     1441 tcctctcccc agccaaagaa gaaaccactg gatggagaat atttcaccct tcagatccgt
     1501 gggcgtgagc gcttcgagat gttccgagag ctgaatgagg ccttggaact caaggatgcc
     1561 caggctggga aggagccagg ggggagcagg gctcactcca gccacctgaa gtccaaaaag
     1621 ggtcagtcta cctcccgcca taaaaaactc atgttcaaga cagaagggcc cgactcagac
     1681 gtctaggtac caagctgatc cgcccctctc cctccccccc ccctaacgtt actggccgaa
     1741 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     1801 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     1861 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     1921 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     1981 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     2041 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     2101 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     2161 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     2221 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     2281 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2341 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2401 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2461 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2521 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2581 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2641 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2701 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2761 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2821 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2881 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2941 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaag cggccgcgac
     3001 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3061 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3121 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3181 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3241 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3301 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3361 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3421 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3481 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3541 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3601 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3661 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3721 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3781 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3841 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4021 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4081 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4141 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4201 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4261 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4321 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4381 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4441 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4501 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4561 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4621 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4681 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4741 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4801 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4861 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4921 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4981 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5041 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5101 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5161 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5221 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5281 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5341 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5401 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5461 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5521 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5581 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5641 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5701 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5761 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6181 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6241 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6301 ataaccgtat taccgccatg cat
//
