LOCUS       sequence-358823-        6836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of C-terminal BirA-tagged TARG1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6836)
  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 6836)
  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-TARG1
FEATURES             Location/Qualifiers
     source          1..6836
                     /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(1395..1742,1743..1745,1746..2354)
                     /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             2358..2384
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2415..2432)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2421..2645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2691..3119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2778..2797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2988..3009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3128..3148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3133..3462
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3313..3448
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3375..3394
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3529..4323
                     /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(3583..3602)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4193..4212
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4497..4618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4534..4553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4588..4607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4667..4683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4667..4683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4680..4702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4691..4707
                     /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(4715..4721,4722..4739,4740..4745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4760..4781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4898..4915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5069..5657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5149..5168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5828..6619,6620..6688))
                     /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     6451..6470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6689..6793)
                     /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 tcatggccag cagccttaat gaagatccag
      961 aaggaagcag aatcacttat gtgaaaggag acctttttgc atgcccgaaa acagactctt
     1021 tagcccactg tatcagtgag gattgtcgca tgggcgctgg gatagctgtc ctctttaaga
     1081 agaaatttgg aggggtgcaa gaacttttaa atcaacaaaa gaaatctgga gaagtggctg
     1141 ttctgaagag agatgggcga tatatatatt acttgattac aaagaaaagg gcttcgcaca
     1201 agccaactta tgaaaactta cagaagagtt tagaggcaat gaagtctcat tgtctgaaga
     1261 atggagtcac tgacctctcc atgcccagga ttggatgtgg tcttgatcgt ctgcaatggg
     1321 aaaatgtatc tgcgatgatc gaggaggtat ttgaggcaac agacatcaaa attactgtgt
     1381 acacactcgg atccaaggac aacaccgtgc ccctgaagct gatcgccctg ctggccaacg
     1441 gcgagttcca ctctggcgag cagctgggag agaccctggg aatgagcaga gccgccatca
     1501 acaagcacat ccagacactg agagactggg gagtggacgt gttcaccgtg cctggcaagg
     1561 gctacagcct gcctgagcct atccagctgc tgaacgccaa gcagatcctg ggacagctgg
     1621 atggcggaag cgtggccgtg ctgcctgtga tcgactccac caatcagtac ctgctggaca
     1681 gaatcggaga gctgaagtcc ggcgacgcct gcatcgccga gtaccagcag gctggcagag
     1741 gaggcagagg acggaagtgg ttcagcccat tcggagccaa cctgtacctg tccatgttct
     1801 ggagactgga gcagggacct gctgctgcca tcggactgag tctggtgatc ggaatcgtga
     1861 tggccgaggt gctgagaaag ctgggagccg acaaggtgag agtgaagtgg cctaatgacc
     1921 tgtacctcca ggaccgcaag ctggctggca tcctggtgga gctgacaggc aagacaggcg
     1981 atgccgctca gatcgtgatc ggagccggaa tcaacatggc catgagaaga gtggaggaga
     2041 gcgtggtgaa ccagggctgg atcaccctgc aggaggctgg catcaacctg gaccggaaca
     2101 ccctggccgc catgctgatc agagagctga gagccgctct ggagctgttc gagcaggagg
     2161 gactggctcc ttacctgagc agatgggaga agctggacaa cttcatcaac agacctgtga
     2221 agctgatcat cggcgacaag gaaatcttcg gcatctccag aggaatcgac aagcagggag
     2281 ctctgctgct ggagcaggac ggaatcatca agccctggat gggcggagaa atctccctga
     2341 gaagcgcaga gaaggcttac ccttacgatg taccggatta cgcataggcg gcctaagttt
     2401 aaaccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2461 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2521 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2581 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2641 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2701 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2761 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2821 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2881 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2941 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3001 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3061 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3121 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3181 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3301 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3361 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3421 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3481 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3541 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3601 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3661 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3721 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3781 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3841 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3901 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3961 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4021 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4081 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4141 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4201 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4261 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4321 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4381 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4441 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4501 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4561 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4621 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4681 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4741 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4801 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4861 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4921 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4981 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5041 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5101 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5161 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5221 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5281 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5341 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5401 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5461 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5521 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5581 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6781 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcggga
//
