LOCUS       sequence-367279-        6021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to see protein dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6021)
  AUTHORS   Poon J, Fries A, Wessel GM, Yajima M
  TITLE     Evolutionary modification of AGS protein contributes to formation of
            micromeres in sea urchins.
  JOURNAL   Nat Commun. 2019 Aug 22;10(1):3779. doi: 10.1038/s41467-019-11560-8.
  PUBMED    31439829
REFERENCE   2  (bases 1 to 6021)
  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: "Nat 
            Commun."; date: "2019-08-22"; volume: "10(1):3779. doi"; pages: " 
            10.1038/s41467-019-11560-8"
COMMENT     Sequence Label: membrane-HsPLK1-dead
FEATURES             Location/Qualifiers
     source          1..6021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..67)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(51..67)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(64..86)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    75..91
                     /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(99..105,106..123,124..129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    144..165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(282..299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(453..1041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(533..552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1212..2003,2004..2072))
                     /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     1835..1854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2073..2177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2245..2263
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2301..2323)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2423..2442
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2656..2675
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2965..2983
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2965..2982
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     5'UTR           2995..3037
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             3130..3837
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3275..3293)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3543..3562
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3883..3903
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3916..3960
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     3'UTR           5829..5954
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
ORIGIN
        1 ctgcaggtcg actctagagg atccccgggc gagctcgaat tcgtaatcat gtcatagctg
       61 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      421 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1201 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1261 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1321 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1381 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1441 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1501 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1561 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1621 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1681 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1741 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1801 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1861 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     1921 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     1981 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2041 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2101 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2161 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2221 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     2281 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2341 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2401 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattcg
     2461 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     2521 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     2581 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     2641 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     2701 ggtgatgccg gccacgatgc gtccggcgta gaggatctgg ctagcgatga ccctgctgat
     2761 tggttcgctg accatttccg ggtgcgggac ggcgttacca gaaactcaga aggttcgtcc
     2821 aaccaaaccg actctgacgg cagtttacga gagagatgat agggtctgct tcagtaagcc
     2881 agatgctaca caattaggct tgtacatatt gtcgttagaa cgcggctaca attaatacat
     2941 aaccttatgt atcatacaca tacgatttag gtgacactat agaatacaag cttgcttgtt
     3001 ctttttgcag aagctcagaa taaacgctca actttggcag atctgatcat gccatggggg
     3061 cccatgctgt gctgtatgag aagaaccaaa caggttgaaa agaatgatga cgaccaaaag
     3121 attggcggca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     3181 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     3241 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     3301 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     3361 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     3421 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     3481 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     3541 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     3601 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     3661 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     3721 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     3781 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtcc
     3841 ggaggggatc ggtggagctc caccggtggc ggccgctcta gagagaatct ttattttcag
     3901 ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca catggacagc
     3961 ggaggtggag gtactagtct cgagatgagt gctgcagtga ctgcagggaa gctggcacgg
     4021 gcaccggccg accctgggaa agccggggtc cccggagttg cagctcccgg agctccggcg
     4081 gcggctccac cggcgaaaga gatcccggag gtcctagtgg acccacgcag ccggcggcgc
     4141 tatgtgcggg gccgcttttt gggcaagggc ggctttgcca agtgcttcga gatctcggac
     4201 gcggacacca aggaggtgtt cgcgggcagg attgtgccta agtctctgct gctcaagccg
     4261 caccagaggg agaagatgtc catggaaata tccattcacc gcagcctcgc ccaccagcac
     4321 gtcgtaggat tccacggctt tttcgaggac aacgacttcg tgttcgtggt gttggagctc
     4381 tgccgccgga ggtctctcct ggagctgcac aagaggagga aagccctgac tgagcctgag
     4441 gcccgatact acctacggca aattgtgctt ggctgccagt acctgcaccg aaaccgagtt
     4501 attcatcgag acctcaagct gggcaacctt ttcctgaatg aagatctgga ggtgaaaata
     4561 ggggattttg gactggcaac caaagtcgaa tatgacgggg agaggaagaa gaccctgtgt
     4621 gggactccta attacatagc tcccgaggtg ctgagcaaga aagggcacag tttcgaggtg
     4681 gatgtgtggt ccattgggtg tatcatgtat accttgttag tgggcaaacc accttttgag
     4741 acttcttgcc taaaagagac ctacctccgg atcaagaaga atgaatacag tattcccaag
     4801 cacatcaacc ccgtggccgc ctccctcatc cagaagatgc ttcagacaga tcccactgcc
     4861 cgcccaacca ttaacgagct gcttaatgac gagttcttta cttctggcta tatccctgcc
     4921 cgtctcccca tcacctgcct gaccattcca ccaaggtttt cgattgctcc cagcagcctg
     4981 gaccccagca accggaagcc cctcacagtc ctcaataaag gcttggagaa ccccctgcct
     5041 gagcgtcccc gggaaaaaga agaaccagtg gttcgagaga caggtgaggt ggtcgactgc
     5101 cacctcagtg acatgctgca gcagctgcac agtgtcaatg cctccaagcc ctcggagcgt
     5161 gggctggtca ggcaagagga ggctgaggat cctgcctgca tccccatctt ctgggtcagc
     5221 aagtgggtgg actattcgga caagtacggc cttgggtatc agctctgtga taacagcgtg
     5281 ggggtgctct tcaatgactc aacacgcctc atcctctaca atgatggtga cagcctgcag
     5341 tacatagagc gtgacggcac tgagtcctac ctcaccgtga gttcccatcc caactccttg
     5401 atgaagaaga tcaccctcct taaatatttc cgcaattaca tgagcgagca cttgctgaag
     5461 gcaggtgcca acatcacgcc gcgcgaaggt gatgagctcg cccggctgcc ctacctacgg
     5521 acctggttcc gcacccgcag cgccatcatc ctgcacctca gcaacggcag cgtgcagatc
     5581 aacttcttcc aggatcacac caagctcatc ttgtgcccac tgatggcagc cgtgacctac
     5641 atcgacgaga agcgggactt ccgcacatac cgcctgagtc tcctggagga gtacggctgc
     5701 tgcaaggagc tggccagccg gctccgctac gcccgcacta tggtggacaa gctgctgagc
     5761 tcacgctcgg ccagcaaccg tctcaaggcc tcctaaacta gtgactgact aggatctggt
     5821 taccactaaa ccagcctcaa gaacacccga atggagtctc taagctacat aataccaact
     5881 tacactttac aaaatgttgt cccccaaaat gtagccattc gtatctgctc ctaataaaaa
     5941 gaaagtttct tcacattcta aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6001 aaaaaacccc cccccccccc c
//
