LOCUS       sequence-358813-        7655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of C-terminal BirA-tagged MACROD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7655)
  AUTHORS   Zaja R, Aydin G, Lippok BE, Feederle R, Luscher B, Feijs KLH
  TITLE     Comparative analysis of MACROD1, MACROD2 and TARG1 expression, 
            localisation and interactome.
  JOURNAL   Sci Rep. 2020 May 19;10(1):8286. doi: 10.1038/s41598-020-64623-y.
  PUBMED    32427867
REFERENCE   2  (bases 1 to 7655)
  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: "Sci Rep."; 
            date: "2020-05-19"; volume: "10(1):8286. doi"; pages: " 
            10.1038/s41598-020-64623-y"
COMMENT     Sequence Label: pcDNA3.1-MCS-BirA(R118G)-HA-MACROD2
FEATURES             Location/Qualifiers
     source          1..7655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        223..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     757..777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     851..870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        851..869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2214..2561,2562..2564,2565..3173)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3177..3203
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3234..3251)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3240..3464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3510..3938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3597..3616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3807..3828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3947..3967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3952..4281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4132..4267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4194..4213
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4348..5142
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4402..4421)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5012..5031
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5316..5437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5353..5372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5407..5426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5486..5502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5486..5502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5499..5521)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5510..5526
                     /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(5534..5540,5541..5558,5559..5564))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5579..5600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5717..5734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5888..6476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5968..5987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6647..7438,7439..7507))
                     /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     7270..7289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7508..7612)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      841 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgc ttaaggcctg
      901 ttaaccggtc gtacgtctcc ggattcgaat tcatgtaccc cagcaacaag aagaaaaagg
      961 tgtggagaga ggagaaagaa cgtttattga agatgacctt agaagagaga cgcaaagaat
     1021 acctaagaga ctatattccc ctgaacagca ttctgtcatg gaaggaggag atgaagggca
     1081 agggccaaaa tgatgaagaa aatactcagg aaacatccca ggtgaagaaa agtttgactg
     1141 aaaaagtttc tctctataga ggtgacatca cattgctaga ggtagatgct atagtcaatg
     1201 ccgcaaatgc cagtcttctt ggaggaggag gtgtggatgg ctgtattcat agagcagccg
     1261 gcccctgttt gctagctgaa tgtcgtaacc tgaatggctg tgatactgga catgcaaaaa
     1321 tcacatgtgg ctatgacctt cctgcaaaat atgtcatcca tactgtaggg ccaatagcca
     1381 ggggccatat taatggttcc cacaaggaag accttgcaaa ttgctataaa tcatctctga
     1441 agctcgtgaa agaaaataac atccgatcag ttgcatttcc ctgcatctca acaggcattt
     1501 atggctttcc caacgagcct gctgcagtca ttgccctcaa caccattaag gaatggcttg
     1561 ccaagaatca ccatgaggtg gatcggatca ttttctgtgt cttcttagaa gttgacttca
     1621 aaatctacaa aaagaaaatg aatgagtttt tctccgtaga cgataataat gaagaagaag
     1681 aggatgttga aatgaaagaa gattcagatg agaacggtcc agaggagaag caaagtgtgg
     1741 aagaaatgga agagcagagc caagatgcag atggtgtcaa cactgtcact gtgcccggcc
     1801 ctgcttcaga agaggcagtt gaagactgta aagatgaaga ttttgcaaag gatgaaaata
     1861 ttacaaaagg cggtgaagtg acagatcatt ctgtgcgtga ccaagatcat cccgatggac
     1921 aagagaatga ttcaacgaag aatgaaataa aaattgaaac agaatcgcag agctcatata
     1981 tggaaacaga agaactttca tcaaaccaag aagatgccgt gattgtggag caaccagaag
     2041 tgattccatt aacagaggac caagaagaaa aagaaggtga aaaagctcca ggcgaggaca
     2101 cacctaggat gcctgggaaa agtgaaggct ccagtgacct agaaaatact ccaggtcctg
     2161 atgcaggggc acaagatgaa gcgaaggaac aaagaaatgg aactaaagga tccaaggaca
     2221 acaccgtgcc cctgaagctg atcgccctgc tggccaacgg cgagttccac tctggcgagc
     2281 agctgggaga gaccctggga atgagcagag ccgccatcaa caagcacatc cagacactga
     2341 gagactgggg agtggacgtg ttcaccgtgc ctggcaaggg ctacagcctg cctgagccta
     2401 tccagctgct gaacgccaag cagatcctgg gacagctgga tggcggaagc gtggccgtgc
     2461 tgcctgtgat cgactccacc aatcagtacc tgctggacag aatcggagag ctgaagtccg
     2521 gcgacgcctg catcgccgag taccagcagg ctggcagagg aggcagagga cggaagtggt
     2581 tcagcccatt cggagccaac ctgtacctgt ccatgttctg gagactggag cagggacctg
     2641 ctgctgccat cggactgagt ctggtgatcg gaatcgtgat ggccgaggtg ctgagaaagc
     2701 tgggagccga caaggtgaga gtgaagtggc ctaatgacct gtacctccag gaccgcaagc
     2761 tggctggcat cctggtggag ctgacaggca agacaggcga tgccgctcag atcgtgatcg
     2821 gagccggaat caacatggcc atgagaagag tggaggagag cgtggtgaac cagggctgga
     2881 tcaccctgca ggaggctggc atcaacctgg accggaacac cctggccgcc atgctgatca
     2941 gagagctgag agccgctctg gagctgttcg agcaggaggg actggctcct tacctgagca
     3001 gatgggagaa gctggacaac ttcatcaaca gacctgtgaa gctgatcatc ggcgacaagg
     3061 aaatcttcgg catctccaga ggaatcgaca agcagggagc tctgctgctg gagcaggacg
     3121 gaatcatcaa gccctggatg ggcggagaaa tctccctgag aagcgcagag aaggcttacc
     3181 cttacgatgt accggattac gcataggcgg cctaagttta aaccgctgat cagcctcgac
     3241 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3301 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3361 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3421 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3481 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3541 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3601 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3661 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3721 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3781 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3841 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3901 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3961 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4141 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4201 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4261 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4321 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4381 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4441 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4501 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4561 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4621 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4681 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4741 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4801 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4861 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4921 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4981 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5041 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5101 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5161 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5221 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5281 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5341 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5401 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5461 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5521 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5581 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5641 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5701 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5761 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5821 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5881 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5941 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6001 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6061 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6121 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6181 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6241 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6301 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6361 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6421 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6481 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6541 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6601 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6661 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6721 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6781 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6841 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6901 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6961 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7021 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7081 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7141 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7201 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7261 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7321 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7381 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7441 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7501 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7561 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7621 cccgaaaagt gccacctgac gtcgacggat cggga
//
