LOCUS       sequence-490957-        4599 bp    DNA     circular SYN 28-OCT-2025
DEFINITION  Mutant EGFP expression plasmid for the creation of a heteroduplex 
            DNA repair assay plasmid with pGEM5Z(+)-EGFP (p111). *Please see 
            notes in comments section..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4599)
  AUTHORS   Zhou B, Huang C, Yang J, Lu J, Dong Q, Sun LZ
  TITLE     Preparation of heteroduplex enhanced green fluorescent protein 
            plasmid for in vivo mismatch repair activity assay.
  JOURNAL   Anal Biochem. 2009 May 1;388(1):167-9. doi: 
            10.1016/j.ab.2009.02.020. Epub 2009 Feb 25.
  PUBMED    19248754
REFERENCE   2  (bases 1 to 4599)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ab.2009.02.020"; journalName: "Anal Biochem"; date: 
            "2009-05-1- 1"; volume: "388"; issue: "1"; pages: "167-9"
COMMENT     Sequence Label: p189
FEATURES             Location/Qualifiers
     source          1..4599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      664..673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(715..736)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(976..995)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1323..1344
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1512..1593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1549..1568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1603..1622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     regulatory      1640..1649
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1679..1698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1680..1698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1705..1722)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1705..1721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1714..1736)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1862..2317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1949..1968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2159..2180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2395..2499
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2500..2568,2569..3360)
                     /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(2718..2737)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3531..4119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4020..4039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4273..4290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4407..4428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4443..4448,4449..4466,4467..4473)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4481..4497
                     /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)."
     primer_bind     4486..4508
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4505..4521
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4505..4521
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4539..4557
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4539..4556
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      601 gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatatcccg
      661 gtcgccacca tggtgagcaa tcgcgaggag ctgttcaccg gggtggtgcc catcctggtc
      721 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      781 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      841 tagcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
      901 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      961 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1021 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1081 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1141 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1201 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1261 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1321 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1381 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     1441 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     1501 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1621 tcaatgtatc atcccgcggc catggcggcc gggagcatgc gacgtcgggc ccaattcgcc
     1681 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1741 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1801 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2281 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     2341 cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga
     2401 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2461 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2521 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2581 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2641 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2701 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2761 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2821 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2881 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2941 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3001 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3061 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3121 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3181 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3241 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3301 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3361 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3421 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3481 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3541 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3601 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3661 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3721 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3781 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3841 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3901 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3961 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4021 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4081 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4141 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4201 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4261 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4321 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4381 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4441 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4501 cacacaggaa acagctatga ccatgattac gccaagctat ttaggtgaca ctatagaata
     4561 ctcaagctat gcatccaacg cgttgggagc tctcgcata
//
