LOCUS       sequence-308052-        6838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transfection plasmid for the exogneous expression of C-terminal 
            miniTurbo-tagged C12orf49 for generation of FlpIn cell lines for 
            BioID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  AUTHORS   Aregger M, Lawson KA, Billmann M, Costanzo M, Tong AHY, Chan K, 
            Rahman M, Brown KR, Ross C, Usaj M, Nedyalkova L, Sizova O, Habsid 
            A, Pawling J, Lin ZY, Abdouni H, Wong CJ, Weiss A, Mero P, Dennis 
            JW, Gingras AC, Myers CL, Andrews BJ, Boone C, Moffat J
  TITLE     Systematic mapping of genetic interactions for de novo fatty acid 
            synthesis identifies C12orf49 as a regulator of lipid metabolism.
  JOURNAL   Nat Metab. 2020 Jun;2(6):499-513. doi: 10.1038/s42255-020-0211-z. 
            Epub 2020 Jun 1.
  PUBMED    32694731
REFERENCE   2  (bases 1 to 6838)
  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/s42255-020-0211-z"; journalName: "Nat Metab"; date: 
            "2020-06"; volume: "2"; issue: "6"; pages: "499-513"
COMMENT     Sequence Label: pCDNA5_C12orf49-miniTurbo
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    987..1011
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1627..1651)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1674..1739
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1779..2735
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     primer_bind     complement(2790..2807)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2796..3020
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3153..3172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3304..3351
                     /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             3359..4381
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4511..4632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4548..4567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4602..4621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4681..4697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4681..4697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4694..4716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4705..4721
                     /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(4729..4735,4736..4753,4754..4759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4774..4795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4912..4929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5083..5671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5163..5182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5842..6633,6634..6702))
                     /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"
     promoter        complement(6703..6807)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag ctttagacaa gtttgtacaa aaaagttggc atggtgaacc
     1021 tggcggccat ggtgtggcgc cggcttctgc ggaagaggtg ggtgctcgcc ctggtcttcg
     1081 ggctgtcgct cgtctacttc ctcagcagca ccttcaagca ggaggagagg gcagtgagag
     1141 ataggaatct cctccaggtt catgaccata atcagcccat cccgtggaaa gtgcagttta
     1201 acttgggcaa tagcagtcgt ccgagcaatc agtgccgcaa ctccattcaa gggaagcacc
     1261 tcatcacgga tgaactcggc tacgtttgcg agaggaagga tttgctggta aatggctgct
     1321 gtaatgtcaa cgtccctagc acgaagcagt actgctgtga tggctgctgg cccaacggct
     1381 gctgcagcgc ctatgagtac tgtgtctcct gctgcctgca gcccaacaag caacttctcc
     1441 tggagcgctt cctcaaccgg gcagccgtgg cattccagaa cctcttcatg gcagtcgaag
     1501 atcactttga gttgtgcctg gccaaatgca ggacctcatc tcagagcgtg cagcatgaga
     1561 acacctaccg ggaccccata gcaaagtatt gctatggaga aagcccgccc gagctcttcc
     1621 ccgcttgccc aactttcttg tacaaagtgg ttgattctag aatcgatggt accgactaca
     1681 aggaccacga cggtgactac aaggaccacg acatcgacta caaggacgac gacgacaagg
     1741 ggggaggggg aggatctggg ggagggggag gatctgcgaa agacaatact gtgcctctga
     1801 agctgatcgc tctcctggct aatggcgagt tccatagtgg cgaacagctg ggagaaaccc
     1861 tgggcatgtc cagggccgct atcaacaagc acattcagac tctgcgcgac tggggcgtgg
     1921 acgtgttcac cgtgcccgga aagggctact ctctgcccga gcctatcccg ctgctgaacg
     1981 ctaaacagat tctgggacag ctggacggcg ggagcgtggc agtcctgcct gtggtcgact
     2041 ccaccaatca gtacctgctg gatcgaatcg gcgagctgaa gagtggggat gcttgcattg
     2101 cagaatatca gcaggcaggg agaggaagca gagggaggaa atggttctct ccttttggag
     2161 ctaacctgta cctgagtatg ttttggcgcc tgaagcgggg accagcagca atcggcctgg
     2221 gcccggtcat cggaattgtc atggcagaag cgctgcgaaa gctgggagca gacaaggtgc
     2281 gagtcaaatg gcccaatgac ctgtatctgc aggatagaaa gctggcaggc atcctggtgg
     2341 agctggccgg aataacaggc gatgctgcac agatcgtcat tggcgccggg attaacgtgg
     2401 ctatgaggcg cgtggaggaa agcgtggtca atcagggctg gatcacactg caggaagcag
     2461 ggattaacct ggacaggaat actctggccg ctacgctgat ccgagagctg cgggcagccc
     2521 tggaactgtt cgagcaggaa ggcctggctc catatctgcc acggtgggag aagctggata
     2581 acttcatcaa tagacccgtg aagctgatca ttggggacaa agagattttc gggattagcc
     2641 gggggattga taaacaggga gccctgctgc tggaacagga cggagttatc aaaccctgga
     2701 tgggcggaga aatcagtctg cggtctgccg aaaagtaagg ccggccccta ggctcgagtc
     2761 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2821 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2881 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2941 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3001 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3061 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3121 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3181 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3241 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     3301 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     3361 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3421 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3481 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3541 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3601 gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3661 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3721 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3781 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3841 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3901 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     3961 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4021 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4081 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     4141 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4201 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4261 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4321 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4381 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4441 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4501 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4561 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4621 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4681 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4741 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4801 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
