LOCUS       sequence-134737-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminal domain of Chk1 with C-term GFP fluorescent tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5411)
  AUTHORS   Zhang M, Kothari P, Mullins M, Lampson MA
  TITLE     Regulation of zygotic genome activation and DNA damage checkpoint 
            acquisition at the mid-blastula transition.
  JOURNAL   Cell Cycle. 2014;13(24):3828-38. doi: 10.4161/15384101.2014.967066.
  PUBMED    25558827
REFERENCE   2  (bases 1 to 5411)
  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: "Cell 
            Cycle."; date: "2014"; volume: "13(24):3828-38. doi"; pages: " 
            10.4161/15384101.2014.967066"
COMMENT     Sequence Label: pMZ040
FEATURES             Location/Qualifiers
     source          1..5411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(702..704,705..707,708..1421)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(702..704,705..707,708..1418)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(747..768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1008..1027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1355..1376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1436..1455)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1437..1455)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1460..1594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1484..1503)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1538..1557
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1707..1727)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1707..1725)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1746..1762)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1746..1762)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1759..1781)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1770..1786
                     /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(1794..1800,1801..1818,1819..1824))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1839..1860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1977..1994)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2148..2736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2228..2247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2907..3698,3699..3767))
                     /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     3530..3549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3768..3872)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3898..4353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4035..4056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4247..4266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4427..5411
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca ggaccattgg tttagcagaa
      121 gtttcaaatc agcagtacaa tgtcagggca tcacaccagc agccaagatt caacgggctg
      181 actctgagct gagacgaaaa aacagtgatg accgagcgca gatctccagc tctcagcctg
      241 agccacaggg attatgggag gagaaggagg ttacagttca caccgatgtc agtgtcagct
      301 tttcacaacc gacctgtcct gaccacatgc tgctgggcac acaactgctt ggcacaccgg
      361 gtgcaagcca gagcccctgg cagcgtttgg tgaggagaat gaccaggttc tttactacag
      421 aaaatacaga ggcatcttgc attgctctcc gtgattcttg tattgctttg ggtcacacat
      481 ggaaacagag ctgtaataat caggcgacag tgtctacaat ggatcgtcgc aataataagc
      541 tgatattcat agtgcacttc ctggaaatgg aggaacgtat tttagttgat ttccggctgt
      601 caaaggggga cggtttggaa ttcaagcgaa tattcctgaa actcaaacag aagctgtctg
      661 acattattag caaccaaaag gttttgccat tgattctcga gatggtgagc aagggcgagg
      721 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      781 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      841 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      901 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      961 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1021 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1081 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1141 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1201 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1261 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1321 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1381 ccgccgggat cactctcggc atggacgagc tgtacaagta atctagattc tgcagcccta
     1441 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     1501 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1561 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1621 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     1681 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     1741 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1801 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1861 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1921 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1981 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2041 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2101 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2161 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2221 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2281 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2341 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2401 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2461 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2521 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2581 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2641 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2701 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2761 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2821 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2881 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2941 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3001 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3061 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3121 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3181 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3241 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3301 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3361 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3421 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3481 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3541 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3601 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3661 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3721 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3781 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3841 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     3901 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3961 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4021 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4081 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4141 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4201 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4261 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4321 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     4381 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc atagccaatt
     4441 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
     4501 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
     4561 caatatggcg gacttggcac catgccaatt caatatggcg gacttggcac tgtgccaact
     4621 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
     4681 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
     4741 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
     4801 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
     4861 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     4921 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     4981 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     5041 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     5101 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     5161 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     5221 aggacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     5281 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     5341 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     5401 gtttagggaa c
//
