LOCUS       sequence-268879-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AGS-mCherry mRNA for microinjection for visualization 
            of AGS protein dynamics in S. purpuratus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  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 6065)
  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: pSP64-AGS-mCherry
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           12..54
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             2044..2751
                     /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(2189..2207)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2457..2476
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2830..2850
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2863..2907
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     3'UTR           2952..3077
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(3133..3149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3146..3168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3157..3173
                     /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(3181..3187,3188..3205,3206..3211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3226..3247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3364..3381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3535..4123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3615..3634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4294..5085,5086..5154))
                     /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     4917..4936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5155..5259)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5327..5345
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5383..5405)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5505..5524
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5738..5757
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6047..6065
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6047..6064
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atacaagctt gcttgttctt tttgcagaag ctcagaataa acgctcaact ttggcagatc
       61 tagatgcaga gcgaggcctc ctgtatggag ctggccctcg aaggggagag gttgtgcaag
      121 gcggggaact gccgggaagg ggtacggtac ctggaggccg ccgtcgaggt cggaaccgat
      181 gatcttaaaa cactaagtgc catctacagt cagctaggca atgcctattt ctatttggag
      241 gactatggca aggccctgac ctaccacaag catgatctga ctctagcgac atcgattggt
      301 gatcgactcg gagaagccaa agctagtggt aatattggca atacactgaa agttctagga
      361 aagtttgatg aagccatctg ctgttgtcaa agacatctag atatatcaag agagctctcg
      421 gaaaaggtgg gggaaggcag ggcactgtac aatctaggca atgtgtacca tgccaaagga
      481 aagcatgctg ggcggagcgg gcatcaagat ccgggagact tcccgcaaga ggttacagac
      541 tgccttaagc aagccatcca gttttatgaa gccaacctag caatagtgag ggagcttgga
      601 gacagagcgg cacaaggcag ggcctgtggt aacctaggca acacacatta tcttctgggc
      661 aacttcgaga cggctataca attccacaca gagcgtcttg ctatagcaaa ggaatttgga
      721 gacaaagcag cagagaggcg agcttacagc aacctaggaa acgcatgtgt cttcatggta
      781 cagtttgaag tggctgccaa atattacaag aaaagtttgc acattgcaag gcagctcggc
      841 gagttggcca tggaggctca ggcgtgctac agtctaggga acacgtatac cttattacgg
      901 gagtatgaga aggccgtgga atatcacagc aggcacatgg agatagctca acaacttaac
      961 gacagagttg gggaaggacg ggcatgctgg agtttaggga acgcccatac gtcattaggt
     1021 aaccacgaga aggctcttca ctatgcaact cttcaccttc aaatatcaag agaggttgga
     1081 gataggacag gagaggtgac agccaaaatg aacttgcaag atttacagac cctctttggt
     1141 ataagtacag ctgatcttag tgaagtcagt acaacagttc aaacacctct acaagaatcc
     1201 aaaggtgcta gaccccgaag aagaagaagt atggaaaatt tggaactagt tgccatgaca
     1261 ccagaaaaga aaacagaaat cgtccaggtc cccaaaagga aaattagacc ggggagtaag
     1321 ctcaaattaa agaatggcgg agaatccaaa gataagcaga gtcataagga ttccaaggag
     1381 tcagttagca gtgccaagaa tagtgtgccc gccccgtcca aagctaaatc acagacctca
     1441 gcacggaagg gaggagaacc tatacagatg ctggacgacg ctgataactt ctttgaggct
     1501 ctcagccgtt tccagagcaa cagaatggac gagcagcgat gctcgttcgg cagactgcag
     1561 aagaagctgg cagatgagga gggaaacgga ttaccagaga aggaagaaca cttagatgag
     1621 atagccaagc ttcagggcag ccgtctcaat gaacagaggg cattctctgt caaacggttg
     1681 ccagggctac ccggtctcag ggccaatgaa gacgttgtag ggaagctgtt agccaaagga
     1741 gaacgagccg agccagatga tgatttcttt gagatgatca ccagatgcca gggagcacgg
     1801 atagaagacc aacgtagcac attacctata caggccccag cccccaccgt cccggatgag
     1861 gacttcttca gtctgatcca gcgcatccag tccaagcgca tcgaggagca gcgatccata
     1921 gcaccgtggg aaaagggttc tggtacatcc tgcgtgtgtt tctatgacta tgacacaagc
     1981 cgcactgaag gaactgcatg ctgcgtctac agcagacctg ttcaagaatc tcacctaggg
     2041 cccatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2101 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2161 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2221 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2281 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2341 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2401 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2461 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2521 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2581 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2641 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2701 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtccggaggg
     2761 gatcggtgga gctccaccgg tcgagctcaa gcttcgaatt ctgcagtcga cggtagcggc
     2821 cgctctagag agaatcttta ttttcagggc gccgccaaat tcaaagaaac cgctgctgct
     2881 aaattcgaac gccagcacat ggacagcgga ggtggaggta ctagtgactg actaggatct
     2941 ggttaccact aaaccagcct caagaacacc cgaatggagt ctctaagcta cataatacca
     3001 acttacactt tacaaaatgt tgtcccccaa aatgtagcca ttcgtatctg ctcctaataa
     3061 aaagaaagtt tcttcacccc ccctgcaggt cgactctaga ggatccccgg gcgagctcga
     3121 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5281 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5341 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5401 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5461 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5521 tgtactgaga gtgcaccatt cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     5581 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     5641 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     5701 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5761 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     5821 ggctagcgat gaccctgctg attggttcgc tgaccatttc cgggtgcggg acggcgttac
     5881 cagaaactca gaaggttcgt ccaaccaaac cgactctgac ggcagtttac gagagagatg
     5941 atagggtctg cttcagtaag ccagatgcta cacaattagg cttgtacata ttgtcgttag
     6001 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     6061 ataga
//
