LOCUS       sequence-439899-        5143 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of FLAG-tagged human p31(comet).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5143)
  AUTHORS   Chuah YH, Tay EXY, Grinchuk OV, Yoon J, Feng J, Kannan S, Robert M, 
            Jakhar R, Liang Y, Lee BWL, Wang LC, Lim YT, Zhao T, Sobota RM, Lu 
            G, Low BC, Crasta KC, Verma CS, Lin Z, Ong DST
  TITLE     CAMK2D serves as a molecular scaffold for RNF8-MAD2 complex to 
            induce mitotic checkpoint in glioma.
  JOURNAL   Cell Death Differ. 2023 Aug;30(8):1973-1987. doi: 
            10.1038/s41418-023-01192-3. Epub 2023 Jul 19.
  PUBMED    37468549
REFERENCE   2  (bases 1 to 5143)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41418-023-01192-3"; journalName: "Cell Death Differ"; 
            date: "2023-08"; volume: "30"; issue: "8"; pages: "1973-1987"
COMMENT     Sequence Label: pCMV-FLAG-p31(comet)
FEATURES             Location/Qualifiers
     source          1..5143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(679..695)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(680..696)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(751..770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(752..770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1044..1165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1081..1100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1135..1154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1172..1627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1309..1330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1521..1540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1654..1758
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1760..2117
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1760..1951
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1783..1803)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1968..2103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2152..2946
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2152..2946
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2206..2225)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2816..2835
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3134..3153)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3178..3225
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3554..4142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4043..4062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4317..4620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4621..4824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4775..4795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        4870..4888
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      4923..4932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4932..4955
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 agactgcatg gtaccagtgg tgtttcctgg gcctgtgagc caggaaggct gctgtcagtt
       61 tacttgtgaa cttctaaagc atatcatgta tcaacgccag cagctccctc tgccctatga
      121 acagcttaag cacttttacc gaaaaccttc tccccaggca gaggagatgc tgaagaagaa
      181 acctcgggcc accactgagg tgagcagcag gaaatgccaa caagccctgg cagaactgga
      241 gagtgtcctc agccacctgg aggacttctt tgcacggaca ctagtaccgc gagtgctgat
      301 tctccttggg ggcaatgccc taagccccaa ggagttctat gaactcgact tgtctctgct
      361 ggccccctac agcgtggacc agagcctgag cacagcagct tgtttgcgcc gtctcttccg
      421 agccatattc atggctgatg cctttagcga gcttcaggct cctccactca tgggcaccgt
      481 cgtcatggca cagggacacc gcaactgtgg agaagattgg tttcgaccca agctcaacta
      541 tcgagtgccc agccggggcc ataaactgac tgtgaccctg tcatgtggca gaccttccat
      601 ccgaaccacg gcttgggaag actacatttg gttccaggca ccagtgacat ttaaaggctt
      661 ccgcgagtga aagcttatcg ataccgtcga cctcgagggg gggcccggta ccttaattaa
      721 ttaaggtacc aggtaagtgt acccaattcg ccctatagtg agtcgtatta caattcactc
      781 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggagatccaa tttttaagtg
      841 tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcac agtcccaagg
      901 ctcatttcag gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat
      961 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1021 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1081 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1141 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     1201 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1261 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     1321 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1381 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     1441 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1501 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     1561 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     1621 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1681 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1741 tattgaaaaa ggaagaatcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     1801 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1861 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1921 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1981 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2041 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2101 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     2161 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2221 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2281 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     2341 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2401 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2461 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     2521 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     2581 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agaacatcag
     2641 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     2701 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     2761 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     2821 gctacccgtg atattgctga agaacttggc ggcgaatggg ctgaccgctt cctcgtgctt
     2881 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     2941 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3001 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3061 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     3121 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3181 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     3241 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     3301 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     3361 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     3421 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3481 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3541 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3601 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3661 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3721 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3781 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3841 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3901 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3961 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4021 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4081 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4141 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4201 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     4261 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4321 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4381 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4441 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4501 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4561 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4621 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     4681 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     4741 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     4801 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgattacgc
     4861 caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg
     4921 ccgccaccat ggattacaag gatgacgacg ataagagccc gggcggatcc cccgggctgc
     4981 aggaattcat ggcggcgccg gaggcggagg ttctgtcctc agccgcagtc cctgatttgg
     5041 agtggtatga gaagtccgaa gaaactcacg cctcccagat agaactactt gagacaagct
     5101 ctacgcagga acctctcaac gcttcggagg ccttttgccc aag
//
