LOCUS       sequence-441735-        7758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MAC3-tagged CCDC22(Y557C) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7758)
  AUTHORS   Laulumaa S, Kumpula EP, Huiskonen JT, Varjosalo M
  TITLE     Structure and interactions of the endogenous human Commander 
            complex.
  JOURNAL   Nat Struct Mol Biol. 2024 Jun;31(6):925-938. doi: 
            10.1038/s41594-024-01246-1. Epub 2024 Mar 8.
  PUBMED    38459129
REFERENCE   2  (bases 1 to 7758)
  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"; doi: 
            "10.1038/s41594-024-01246-1"; journalName: "Nat Struct Mol Biol"; 
            date: "2024-06"; volume: "31"; issue: "6"; pages: "925-938"
COMMENT     Sequence Label: MAC3_C_CCDC22(Y557C)
FEATURES             Location/Qualifiers
     source          1..7758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    802..826
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2708..2732)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2779..2802
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2839..2862
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2893..2919
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2893..2919
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3476..3493)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3482..3706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3839..3858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3990..4037
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4045..5067
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4045..5067
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5197..5318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5234..5253)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5288..5307
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5367..5383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5367..5383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5380..5402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5391..5407
                     /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(5415..5421,5422..5439,5440..5445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5460..5481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5598..5615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5769..6357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5849..5868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6528..7319,7320..7388))
                     /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     7151..7170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7389..7493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7568..7587)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt
      781 ggtggaattc tgcagatatc aacaagtttg tacaaaaaag ttggcatgga ggaggcggac
      841 cgaatcctca tccattcgct gcgccaggcc ggcacggcag ttcctccaga tgtgcagacc
      901 ttgcgcgcct tcaccactga gctggttgta gaggctgtgg tccgctgcct gcgtgtgatc
      961 aaccctgcgg tgggctctgg cctcagccct ctgctgcctc ttgccatgtc tgcccggttc
     1021 cgcctggcca tgagcctggc tcaggcctgc atggacctgg gctatccctt ggagcttggc
     1081 tatcagaact tcctctaccc cagtgagcct gacctccgag acctgcttct cttcttggct
     1141 gagcgtctgc ccaccgatgc ctctgaggat gcagaccagc ctgcaggtga ctcagctatt
     1201 ctcctccggg ccattgggag ccaaattcgg gaccagctgg cactgccttg ggtcccgccc
     1261 caccttcgca ctcccaagct gcagcacctc cagggctcgg ccctccagaa gcctttccat
     1321 gccagcaggc tggtcgtgcc agaattgagt tccagaggtg agccacggga gttccaggcg
     1381 agtcccctgc tgcttccagt ccctacccag gtgcctcagc ctgttggaag ggtggcctcg
     1441 ctcctcgaac accatgccct gcagctctgc cagcagacgg gccgggaccg gccaggggat
     1501 gaggactggg tccaccggac atcccgcctc ccaccccagg aggacacacg ggctcagcgg
     1561 cagcggctgc agaagcaact gactgagcat ctgcgccaaa gctggggcct gcttggggcc
     1621 cccatacaag cccgggacct gggagaactg ctgcaggcct ggggtgctgg ggccaagact
     1681 ggtgctccta agggctcccg cttcacgcac tcagagaagt tcaccttcca tctggagccc
     1741 caggcccagg ccactcaggt gtcagatgtg ccagccacct cccggcggcc tgaacaggtc
     1801 acgtgggcag ctcaggaaca ggagctcgag tcccttcggg agcagctgga aggagtgaac
     1861 cgcagcattg aggaggttga ggccgacatg aagaccctgg gcgtcagctt tgtgcaggca
     1921 gagtctgagt gccggcacag caagctcagt acagcagagc gtgagcaggc cctgcgcctg
     1981 aagagccgcg cggtggagct gctgcccgat gggactgcca accttgccaa gctgcagctt
     2041 gtggtggaga atagtgccca gcgggtcatc cacttggcgg gtcagtggga gaagcaccgg
     2101 gtcccactcc tcgctgagta ccgccacctc cgaaagctgc aggattgcag agagctggaa
     2161 tcttctcgac ggctggcaga gatccaagaa ctgcaccaga gtgtccgggc ggctgctgaa
     2221 gaggcccgca ggaaggagga ggtctataag cagctgatgt cagagctgga gactctgccc
     2281 agagatgtgt cccggctggc ctacacccag cgcatcctgg agatcgtggg caacatccgg
     2341 aagcagaagg aagagatcac caagatcttg tctgatacga aggagcttca gaaggaaatc
     2401 aactccctat ctgggaagct ggaccggacg tttgcggtga ctgatgagct tgtgttcaag
     2461 gatgccaaga aggacgatgc tgttcggaag gcctgtaagt atctagctgc tctgcacgag
     2521 aactgcagcc agctcatcca gaccatcgag gacacaggca ccatcatgcg ggaggttcga
     2581 gacctcgagg agcagatcga gacagagctg ggcaagaaga ccctcagcaa cctggagaag
     2641 atccgggagg actaccgagc cctccgccag gagaacgctg gcctcctagg ccgggtccgg
     2701 gaggcctgcc caactttctt gtacaaagtg gttgatatca aggccgccga catcaccagc
     2761 ctgtacaaga aagtggggtg gagccatccc cagttcgaga agggcggagg atctggcggc
     2821 ggaagcggcg gaggaagctg gtcccaccct cagtttgaga aaggcaccga gctgggcagc
     2881 accatggcca gctaccccta cgacgtgccc gactacgccg acttcaagaa cctgatctgg
     2941 ctgaaggagg tggacagcac ccaggagaga ctgaaggagt ggaacgtgtc ctacggcacc
     3001 gccctggtgg ccgacagaca gaccaagggc agaggcggcc cgggcagaaa gtggctgagc
     3061 caggagggcg gcctgtactt cagcttcctg ctgaacccca aggagttcga gaacctgctg
     3121 cagctgcccc tggtgctggg cctgagcgtg agcgaggccc tggaggagat caccgagatc
     3181 cccttcagcc tgaagtggcc caacgacgtg tacttccagg agaagaaggt gagcggcgtg
     3241 ctgtgcgagc tgagcaagga caagctgatc gtgggcatcg gcatcaacgt gaaccagaga
     3301 gagatccccg aggagatcaa ggacagagcc accaccctgt acgagatcac cggcaaggac
     3361 tgggacagaa aggaggtgct gctgaaggtg ctgaagagaa tcagcgagaa cctgaagaag
     3421 ttcaaggaga agtgatgact cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3481 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3541 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3601 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3661 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3721 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3781 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3841 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3901 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3961 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta
     4021 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     4081 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     4141 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     4201 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     4261 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     4321 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     4381 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     4441 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     4501 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     4561 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     4621 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     4681 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     4741 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     4801 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     4861 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     4921 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     4981 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     5041 agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc
     5101 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5161 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5221 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5281 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5341 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5401 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5461 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5521 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5581 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5641 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5701 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5761 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5821 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5881 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5941 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6001 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6061 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7501 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     7561 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     7621 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7681 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     7741 gggccagata tacgcgtt
//
