pCMV-RT-MLH1dn-EGFP

LOCUS       pCMV-RT-MLH1dn-EGFP             8688 bp DNA     circular SYN 06-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8688)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..466
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        467..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     621..641
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     712..731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        712..730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     771..828
                     /label=OL1012
     primer_bind     complement(774..830)
                     /label=OL1013
     primer_bind     complement(778..800)
                     /label=OL1036
     CDS             780..800
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     781..826
                     /label=OL1037
     CDS             800
                     /label=SGGSx2-sv40NLS-SGGSx2
                     /note="/created_by=chenp"
     CDS             801..2831
                     /label=RT codon optimized Genscript
                     /note="/created_by=jessiedavis"
     CDS             2877..2897
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2907..2933
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2943..2999
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     2943..2967
                     /label=OL899
     primer_bind     2943..2966
                     /label=PE001
     primer_bind     complement(2981..3000)
                     /label=PE002
     CDS             3000..5255
                     /label=MLH1 del754-756 genscript codon
                     /note="/created_by=chenp"
                     /note="/modified_by=chenp"
     primer_bind     5226..5280
                     /label=OL899
     primer_bind     5255..5279
                     /label=PE001
     CDS             5256..5312
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(5294..5320)
                     /label=PE002
     primer_bind     5304..5330
                     /label=PE003
     CDS             5313..6032
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6008..6062)
                     /label=OL900
     primer_bind     complement(6009..6033)
                     /label=PE004
     primer_bind     complement(6015..6057)
                     /label=OL1038
     primer_bind     6029..6062
                     /label=OL1014
     primer_bind     6038..6061
                     /label=OL1035
     CDS             6038..6055
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6078..6095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6084..6308
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal
                     牛生长激素聚腺苷酸化信号
                     牛生长激素（BGH）多聚腺苷酸化信号能够保证高效的转录终止和mRNA的poly A结构"
     primer_bind     complement(6379..6395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6379..6395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6392..6414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6403..6419
                     /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(6427..6457)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6472..6493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6610..6627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6781..7369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6861..6880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7540..8400)
                     /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     8163..8182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8401..8505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8584..8603)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
       61 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
      121 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      181 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      241 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      301 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      361 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      421 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      481 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      541 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      601 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      661 aagcagagct ggtttagtga accgtcagat ccgctagaga tccgcggccg ctaatacgac
      721 tcactatagg gagagccgcc accatgaaac ggacagccga cggaagcgag ttcgagtcac
      781 caaagaagaa gcggaaagtc accctgaaca ttgaagacga gtatagactg catgaaacaa
      841 gcaaggaacc cgacgtgtcc ctgggctcca cctggctgtc cgactttccc caggcctggg
      901 ccgagacagg aggaatgggc ctggccgtgc ggcaggcacc cctgatcatc cctctgaagg
      961 ccacctctac acccgtgagc atcaagcagt accctatgtc tcaggaggcc agactgggca
     1021 tcaagcctca catccagagg ctgctggacc agggcatcct ggtgccatgc cagagcccct
     1081 ggaacacacc actgctgccc gtgaagaagc caggcaccaa tgactataga cccgtgcagg
     1141 atctgagaga ggtgaacaag agggtggagg atatccaccc caccgtgccc aacccttaca
     1201 atctgctgtc cggcctgccc ccttctcacc agtggtatac agtgctggac ctgaaggatg
     1261 ccttcttttg tctgagactg caccctacca gccagccact gttcgccttt gagtggaggg
     1321 accctgagat gggcatctct ggccagctga cctggacacg cctgcctcag ggcttcaaga
     1381 atagcccaac actgtttaac gaggccctgc accgcgacct ggcagatttc cggatccagc
     1441 acccagatct gatcctgctg cagtacgtgg acgatctgct gctggccgcc accagcgagc
     1501 tggattgcca gcagggaaca cgcgccctgc tgcagaccct gggaaacctg ggatataggg
     1561 catccgccaa gaaggcccag atctgtcaga agcaggtgaa gtacctgggc tatctgctga
     1621 aggagggcca gagatggctg acagaggcca ggaaggagac agtgatgggc cagccaacac
     1681 ccaagacccc aagacagctg agggagttcc tgggcaaagc aggattttgc aggctgttca
     1741 tcccaggatt cgcagagatg gcagcacctc tgtacccact gaccaagccg ggcaccctgt
     1801 ttaattgggg ccctgaccag cagaaggcct atcaggagat caagcaggcc ctgctgacag
     1861 caccagccct gggcctgcca gacctgacca agcctttcga gctgtttgtg gatgagaagc
     1921 agggctacgc caagggcgtg ctgacccaga agctgggacc atggagacgg cccgtggcct
     1981 atctgtccaa gaagctggac ccagtggcag caggatggcc accatgcctg aggatggtgg
     2041 cagcaatcgc cgtgctgaca aaggatgccg gcaagctgac catgggacag ccactggtca
     2101 tcctggcacc acacgcagtg gaggccctgg tgaagcagcc tccagatcgc tggctgtcta
     2161 acgcccggat gacacactac caggccctgc tgctggacac cgatcgcgtg cagtttggcc
     2221 ctgtggtggc cctgaatcca gccaccctgc tgcctctgcc agaggagggc ctgcagcaca
     2281 actgtctgga catcctggca gaggcacacg gaacaaggcc agacctgacc gatcagcccc
     2341 tgcctgacgc cgatcacaca tggtataccg atggaagctc cctgctgcag gagggccaga
     2401 ggaaggcagg agcagcagtg accacagaga cagaagtgat ctgggccaag gccctgccag
     2461 caggcacatc cgcccagcgg gccgagctga tcgccctgac ccaggccctg aagatggccg
     2521 agggcaagaa gctgaacgtg tacacagact ccagatatgc cttcgccacc gcacacatcc
     2581 acggagagat ctacaggcgc cggggctggc tgacctctga gggcaaggag atcaagaaca
     2641 aggatgagat cctggccctg ctgaaggccc tgtttctgcc caagcggctg agcatcatcc
     2701 actgtcctgg acaccagaag ggacactccg ccgaggcaag gggcaatcgg atggccgacc
     2761 aggccgccag aaaggctgct attactgaaa ctcccgacac ttccactctg ctgattgaaa
     2821 actcctcccc ttctggcggc tcaaaaagaa ccgccgacgg cagcgaattc gagtctccca
     2881 agaagaagag gaaagtcggc tctggccctg ccgctaagag agtgaagctg gacggaagcg
     2941 gagctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta
     3001 gcttcgttgc tggagtcatc cggagactgg acgagacagt ggtgaacaga attgccgccg
     3061 gcgaggtgat ccagagacct gccaatgcaa taaaggagat gatcgagaac tgtctggacg
     3121 ccaagtccac aagcattcag gtgatcgtga aggagggcgg actgaagctg atccagatcc
     3181 aagacaacgg cacaggcatc agaaaggaag atctggacat cgtgtgtgaa cggttcacca
     3241 catctaagct gcagtctttt gaggatctgg cctctatcag tacctacggc ttcagaggcg
     3301 aggccctggc cagcatcagc cacgtggccc atgtgaccat caccaccaaa accgccgacg
     3361 gcaaatgcgc ttatcgcgct agctacagcg acggcaagct gaaagccccg ccaaagcctt
     3421 gcgccggcaa ccagggtaca cagataacag tggaggatct gttctacaac atcgccaccc
     3481 ggagaaaggc cctgaaaaat cccagcgagg agtacggcaa gatcctggaa gtcgtcggca
     3541 gatactccgt gcacaacgcc ggaatcagct ttagcgtaaa gaagcaggga gaaaccgtgg
     3601 ccgatgtgcg caccctgcca aatgccagca ccgtggataa catcagaagc attttcggaa
     3661 atgccgtgtc cagagaactg atcgagatcg gctgcgaaga taagaccctg gcttttaaga
     3721 tgaacggcta catcagcaac gccaattact ctgtgaagaa gtgcatcttt cttctgttca
     3781 tcaaccacag actggtggaa agcaccagcc tgcggaaagc catcgagaca gtgtacgccg
     3841 cctacctgcc taagaacacc caccccttcc tgtacctgag cctcgagatc agccctcaga
     3901 acgtggacgt caatgtgcat cctacaaagc acgaggtgca cttcctgcac gaggaaagca
     3961 tcctggaaag agtgcagcag cacattgaga gcaagctgct gggctctaac agcagcagaa
     4021 tgtacttcac acagaccctg ctgcctggcc tggccggccc ctcaggcgaa atggttaagt
     4081 ccacaacctc tctgacctca tccagcacca gcggttcttc cgataaggtg tacgcccacc
     4141 agatggtgcg gaccgactct cgggagcaga agctggacgc ctttctgcaa cctctgagca
     4201 aacctctgag ctctcagcct caggccatcg tgaccgagga caagacagat atctcctccg
     4261 gccgtgccag acagcaggac gaagaaatgc tcgagctgcc agctcctgcc gaggtggccg
     4321 ccaagaacca gagcctggag ggagatacca caaagggcac cagcgaaatg agcgagaagc
     4381 ggggccctac ctccagcaac cccagaaaac ggcaccggga ggacagcgac gtggaaatgg
     4441 tggaggacga cagcagaaag gaaatgacag ccgcttgtac ccctagaaga agaatcatca
     4501 acctgacctc cgtgctgagc ctgcaggagg agatcaacga gcagggccac gaggtgctga
     4561 gagagatgct gcacaatcac agcttcgtgg gctgcgtgaa ccctcaatgg gccctggctc
     4621 agcatcaaac aaagctgtac ctgctgaaca ccaccaagct gagcgaagag ctgttctacc
     4681 agatcctcat ctacgacttc gccaacttcg gcgtgctacg cctgagcgag cccgcccctc
     4741 tgtttgacct ggccatgctg gctctggata gcccagaaag cggctggaca gaagaggacg
     4801 gacctaaaga ggggctggct gaatacatcg tggagttcct gaagaaaaag gccgagatgc
     4861 tggccgacta cttttctctg gaaatcgacg aggaaggcaa cctgatcggc ctgcctctgc
     4921 tgatcgataa ctacgtgcct cccctggaag gcctgcccat cttcatcctg agactggcta
     4981 cagaggtgaa ctgggacgag gaaaaggaat gcttcgagtc tctgagcaag gagtgcgcca
     5041 tgttctatag catcagaaaa cagtacatct ctgaagagag cactctgtct ggccagcaga
     5101 gtgaagtgcc cggaagcatc cccaacagct ggaagtggac cgtggaacac atcgtgtaca
     5161 aggccctgcg gagccacatt ctccctccta agcacttcac cgaggacggc aacatcctgc
     5221 agctggccaa cctgcccgac ctttataagg ttttcgctac taacttcagc ctgctgaagc
     5281 aggctggaga cgtggaggag aaccctggac ctatggtgag caagggcgag gagctgttca
     5341 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     5401 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     5461 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     5521 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     5581 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     5641 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     5701 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     5761 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     5821 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     5881 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     5941 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     6001 tcactctcgg catggacgag ctgtacaagt aaccggtcat catcaccatc accattgagt
     6061 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     6121 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     6181 tgagaaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     6241 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     6301 ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga cctctagcta
     6361 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6421 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag
     6481 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6541 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6601 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6661 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6721 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6781 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6841 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6901 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6961 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7021 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7081 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7141 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7201 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7261 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7381 gatcttttct acggggtctg acactcagtg gaacgaaaac tcacgttaag ggattttggt
     7441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8521 agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag ggtctactct
     8581 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     8641 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggca
//