LOCUS       sequence-128621-        5118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal kinase 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 5118)
  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 5118)
  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: pMZ039
FEATURES             Location/Qualifiers
     source          1..5118
                     /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(409..411,412..414,415..1128)
                     /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(409..411,412..414,415..1125)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(454..475)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(715..734)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1062..1083
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1143..1162)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1144..1162)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1167..1301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1191..1210)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1245..1264
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1414..1434)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1414..1432)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1453..1469)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1453..1469)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1466..1488)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1477..1493
                     /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(1501..1507,1508..1525,1526..1531))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1546..1567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1684..1701)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1855..2443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1935..1954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2614..3405,3406..3474))
                     /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     3237..3256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3475..3579)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3605..4060)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3742..3763)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3954..3973
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4134..5118
                     /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 ggaccatggc tgtgcctttt
      121 gttaaagact gggatgtggt acaaactctt ggagagggag catatggaga ggtgcgactg
      181 ctggtcaaca agaaaacaga agaggctgtg gcggtgaaag ttgtggacat ggcaaaagcc
      241 aaggattgca tcgagaatgt gaagaaggag gtctgcatat gcaagatgct ttcacacccc
      301 aacattgtac gtttctttgg gcacagtgtt gggattacac atcgtgacat aaagcctgag
      361 aatattcttc ttgatgataa agataatctg aagatctctg acctcgagat ggtgagcaag
      421 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      481 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      541 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      601 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      661 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      721 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      781 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      841 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
      901 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      961 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1021 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1081 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatc tagattctgc
     1141 agccctatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
     1201 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1261 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1321 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc
     1381 caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     1441 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1501 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1561 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1621 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1681 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1741 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1801 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1861 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1921 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1981 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2041 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2101 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2161 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2221 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2281 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2341 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2401 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2461 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2521 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2581 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2641 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2701 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2761 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2821 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2881 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2941 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3001 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3061 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3121 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3181 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3241 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3301 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3361 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3421 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3481 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3541 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3601 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3661 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3721 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3781 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3841 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3901 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3961 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4021 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     4081 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccata
     4141 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
     4201 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
     4261 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ttggcactgt
     4321 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
     4381 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
     4441 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
     4501 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
     4561 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
     4621 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
     4681 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
     4741 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
     4801 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
     4861 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
     4921 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
     4981 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
     5041 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
     5101 aatgctcgtt tagggaac
//
