LOCUS       sequence-309550-        7419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7419)
  AUTHORS   Liu X, Salokas K, Weldatsadik RG, Gawriyski L, Varjosalo M
  TITLE     Combined proximity labeling and affinity purification-mass 
            spectrometry workflow for mapping and visualizing protein 
            interaction networks.
  JOURNAL   Nat Protoc. 2020 Aug 10. pii: 10.1038/s41596-020-0365-x. doi: 
            10.1038/s41596-020-0365-x.
  PUBMED    32778839
REFERENCE   2  (bases 1 to 7419)
  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 Protoc.
            2020 Aug 10. pii: 10.1038/s41596-020-0365-x. doi: 
            10.1038/s41596-020-0365-x."
COMMENT     Sequence Label: MAC-GPR78
FEATURES             Location/Qualifiers
     source          1..7419
                     /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(1922..1946)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1993..2016
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2053..2076
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2107..2133
                     /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             2107..2133
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2134..2481,2482..2484,2485..3093)
                     /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"
     primer_bind     complement(3137..3154)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3143..3367
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3500..3519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3651..3698
                     /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             3706..4728
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3706..4728
                     /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    4858..4979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4895..4914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4949..4968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5028..5044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5028..5044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5041..5063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5052..5068
                     /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(5076..5082,5083..5100,5101..5106))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5121..5142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5259..5276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5430..6018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5510..5529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6189..6980,6981..7049))
                     /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     6812..6831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7050..7154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7229..7248)
                     /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 caggctccac catggggcca
      841 ggtgaagctc ttcttgctgg cttgctggtt atggttttgg ccgttgccct cctgtctaac
      901 gcacttgtac tgctgtgctg tgcatattca gccgagctga gaactagagc ctcaggcgtg
      961 ctcctggtaa acctcagcct gggacatctg ctgctggcag ccctggatat gcctttcacc
     1021 ctcctggggg ttatgagagg gcggaccccg tctgcccccg gcgcgtgcca ggtgatcggt
     1081 tttctggaca cgttcctcgc ctcaaatgct gccctgagcg ttgctgccct gtcagctgac
     1141 cagtggctgg cggtcggatt tcctcttcgg tacgccggcc gactgaggcc acgctatgcc
     1201 gggctgcttc tcggatgtgc ctgggggcag agcctcgcat tttcaggagc cgccctcggg
     1261 tgcagctggt tgggatattc cagtgcattt gcatcctgta gcctccggct gccccctgag
     1321 ccagagagac ccaggttcgc cgcctttact gccacactcc atgccgtcgg gtttgttctt
     1381 ccactggctg tcctgtgcct taccagcctg caggtacatc gcgtggcgag gagacactgc
     1441 cagcggatgg acaccgttac catgaaagct ctggcactgc tggctgacct ccatccaagt
     1501 gtccgccaac ggtgcctgat tcagcagaag aggcgacgac accgcgctac aagaaaaata
     1561 ggtatcgcca ttgccacttt tctcatctgt tttgcaccat acgtgatgac taggctggct
     1621 gagctggtcc ctttcgttac cgttaacgcc caatggggaa tactctccaa gtgcctcacc
     1681 tacagcaaag ccgtagctga tccctttaca tattccctcc tgcggcgccc gttcaggcag
     1741 gtactggcgg gtatggtcca tcgcttgctg aaacggactc cacggccggc aagcacccat
     1801 gatagttctc tcgatgttgc aggcatggtg caccaactct tgaaacgcac tcctagacct
     1861 gcgagcacac acaacggctc tgtggacact gaaaatgaca gttgcctcca acagactcac
     1921 gacccagctt tcttgtacaa agtggttgat atcaaggccg ccgacatcac cagcctgtac
     1981 aagaaagtgg ggtggagcca tccccagttc gagaagggcg gaggatctgg cggcggaagc
     2041 ggcggaggaa gctggtccca ccctcagttt gagaaaggca ccgagctggg cagcaccatg
     2101 gccagctacc cctacgacgt gcccgactac gccaaggaca acaccgtgcc cctgaagctg
     2161 atcgccctgc tggccaacgg cgagttccac tctggcgagc agctgggcga gacactgggc
     2221 atgagcagag ccgccatcaa caagcacatc cagaccctgc gggactgggg cgtggacgtg
     2281 ttcacagtgc ccggcaaggg ctacagcctg cccgagccca tccagctgct gaacgccaag
     2341 cagatcctgg gccagctgga tggcggatct gtggccgtgc tgcccgtgat cgacagcacc
     2401 aaccagtatc tgctcgaccg gatcggcgag ctgaagtccg gcgacgcctg tatcgccgag
     2461 taccagcagg ccggcagagg cggcaggggc agaaagtggt tcagcccctt cggcgccaac
     2521 ctgtacctga gcatgttctg gagactggaa cagggcccag ccgccgctat tggcctgagc
     2581 ctggtcatcg gcatcgtgat ggccgaggtg ctgcggaagc tgggcgccga caaagtgcgc
     2641 gtgaagtggc ccaacgacct gtatctgcag gaccggaagc tggccggcat cctggtggaa
     2701 ctgaccggca aaacaggcga cgccgcccag atcgtgatcg gcgctggcat caacatggcc
     2761 atgcggcggg tggaagagag cgtggtcaac cagggctgga tcaccctgca ggaagccggc
     2821 atcaacctgg accggaacac cctggccgcc atgctgatca gagagctgag agccgccctg
     2881 gaactgttcg agcaggaagg cctggccccc tacctgagca gatgggagaa gctggacaac
     2941 ttcatcaacc ggcccgtgaa actgatcatc ggcgacaaag agatcttcgg catcagccgg
     3001 ggcatcgaca agcagggggc cctgctcctg gaacaggacg gcatcatcaa gccctggatg
     3061 ggcggcgaga tcagcctgcg gagcgccgag aagtgatgac tcgagtctag agggcccgtt
     3121 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3181 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3241 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3301 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3361 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3421 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3481 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3541 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3601 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta
     3661 ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc
     3721 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
     3781 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     3841 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     3901 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc
     3961 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     4021 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     4081 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     4141 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     4201 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     4261 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     4321 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     4381 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     4441 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     4501 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     4561 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     4621 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     4681 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga
     4741 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4801 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4861 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4921 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4981 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5041 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5101 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5161 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5221 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5281 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5341 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5401 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5461 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5521 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5581 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5641 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5701 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5761 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5821 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5881 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5941 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6001 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6061 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6121 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6181 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6241 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6301 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6361 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6421 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6481 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6541 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6601 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6661 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6721 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6781 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6841 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6901 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6961 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7021 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7081 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7141 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7201 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7261 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7321 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     7381 ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
