LOCUS       sequence-171235-        8290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8290)
  AUTHORS   Chen W, Arroyo JD, Timmons JC, Possemato R, Hahn WC
  TITLE     Cancer-associated PP2A Aalpha subunits induce functional 
            haploinsufficiency and tumorigenicity.
  JOURNAL   Cancer Res. 2005 Sep 15. 65(18):8183-92.
  PUBMED    16166293
REFERENCE   2  (bases 1 to 8290)
  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: "Cancer Res.
            2005 Sep 15. 65(18):8183-92."
COMMENT     Sequence Label: pMIG-Aalpha WT
FEATURES             Location/Qualifiers
     source          1..8290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..23
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(61..83)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     183..202
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     396..418
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     546..565
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             765..1281
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1345..1686
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1753..2169
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2099..2121
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2137..2153
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2185..2208
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4000..4566
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4016..4568
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4160..4177)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4387..4406
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4580..4582,4583..4585,4586..5299)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4625..4646)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4886..4905)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5233..5254
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5449..5963
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6125..6141
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6149..6155,6156..6173,6174..6179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6194..6215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6332..6349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6503..7091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6583..6602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7262..8053,8054..8122))
                     /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     7885..7904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8123..8227)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
       61 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      121 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      181 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
      241 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
      301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      421 ggcgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
      481 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
      541 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
      601 gccacgatgc gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg
      661 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
      721 tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
      781 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac
      841 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa
      901 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
      961 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     1021 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     1081 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     1141 cagtcctccg atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt
     1201 ttgcatccga atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc
     1261 ccacctcggg ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga
     1321 ccaccgaccc ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc
     1381 tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac
     1441 tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac
     1501 cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag
     1561 tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa
     1621 cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt
     1681 ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     1741 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     1801 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     1861 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     1921 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     1981 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     2041 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     2101 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     2161 ctaggcgccg gaattagatc catggactac aaagacgatg acgacaaggc ggcggccgac
     2221 ggcgacgact cgctgtaccc catcgcggtg ctcatagacg aactccgcaa tgaggacgtt
     2281 cagcttcgcc tcaacagcat caagaagctg tccaccatcg ccttggccct tggggttgaa
     2341 aggacccgaa gtgagcttct gcctttcctt acagatacca tctatgatga agatgaggtc
     2401 ctcctggccc tggcagaaca gctgggaacc ttcactaccc tggtgggagg cccagagtac
     2461 gtgcactgcc tgctgccacc gctggagtcg ctggccacag tggaggagac agtggtgcgg
     2521 gacaaggcag tggagtcctt acgggccatc tcacacgagc actcgccctc tgacctggag
     2581 gcgcactttg tgccgctagt gaagcggctg gcgggcggcg actggttcac ctcccgcacc
     2641 tcggcctgcg gcctcttctc cgtctgctac ccccgagtgt ccagtgctgt gaaggcggaa
     2701 cttcgacagt acttccggaa cctgtgctca gatgacaccc ccatggtgcg gcgggccgca
     2761 gcctccaagc tgggggagtt tgccaaggtg ctggagctgg acaacgtcaa gagtgagatc
     2821 atccccatgt tctccaacct ggcctctgac gagcaggact cggtgcggct gctggcggtg
     2881 gaggcgtgcg tgaacatcgc ccagcttctg ccccaggagg atctggaggc cctggtgatg
     2941 cccactctgc gccaggccgc tgaagacaag tcctgggccg tccgctacat ggtggctgac
     3001 aagttcacag agctccagaa agcagtgggg cctgagatca ccaagacaga cctggtccct
     3061 gccttccaga acctgatgaa agactgtgag gccgaggtga gggccgcagc ctcccacaag
     3121 gtcaaagagt tctgtgaaaa cctctcagct gactgtcggg agaatgtgat catgtcccag
     3181 atcttgccct gcatcaagga gctggtgtcc gatgccaacc aacatgtcaa gtctgccctg
     3241 gcctcagtca tcatgggtct ctctcccatc ttgggcaaag acaacaccat cgagcacctc
     3301 ttgcccctct tcctggctca gctgaaggat gagtgccctg aggtacggct gaacatcatc
     3361 tctaacctgg actgtgtgaa cgaggtgatt ggcatccggc agctgtccca gtccctgctc
     3421 cctgccattg tggagctggc tgaggacgcc aagtggcggg tgcggctggc catcattgag
     3481 tacatgcccc tcctggctgg acagctggga gtggagttct ttgatgagaa acttaactcc
     3541 ttgtgcatgg cctggcttgt ggatcatgta tatgccatcc gcgaggcagc caccagcaac
     3601 ctgaagaagc tagtggaaaa gtttgggaag gagtgggccc atgccacaat catccccaag
     3661 gtcttggcca tgtccggaga ccccaactac ctgcaccgca tgactacgct cttctgcatc
     3721 aatgtgctgt ctgaggtctg tgggcaggac atcaccacca agcacatgct acccacggtt
     3781 ctgcgcatgg ctggggaccc ggttgccaat gtccgcttca atgtggccaa gtctctgcag
     3841 aagatagggc ccatcctgga caacagcacc ttgcagagtg aagtcaagcc catcctagag
     3901 aagctgaccc aggaccagga tgtggacgtc aaatactttg cccaggaggc tctgactgtt
     3961 ctgtctctcg cctgagtcga ctcgagcatg cccgggaatt ccgccccccc ccctaacgtt
     4021 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     4081 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     4141 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     4201 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     4261 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     4321 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     4381 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     4441 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     4501 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     4561 tgataatatg gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4621 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg
     4681 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4741 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4801 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4861 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4921 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4981 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     5041 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     5101 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     5161 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     5221 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     5281 ggacgagctg tacaagtaat gaattaatta agaattatca ccgcttctat tcagccagta
     5341 aggcctgtct ttaatggcct ccggcatgag acactttcta gagtcgacct gcagccaagc
     5401 ttatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     5461 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat
     5521 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
     5581 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     5641 ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     5701 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     5761 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
     5821 cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc
     5881 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     5941 acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg
     6001 agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat
     6061 actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt
     6121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6241 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6301 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6361 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6421 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6481 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6541 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6601 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6661 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6721 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6781 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6841 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6901 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6961 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7021 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7081 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7141 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7201 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7261 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7321 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7381 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7441 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7501 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7561 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7621 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7681 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7741 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7801 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7861 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7921 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7981 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8041 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8101 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8161 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8221 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8281 cattaaccta
//
