LOCUS       sequence-333382-        7563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ETS1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7563)
  AUTHORS   Goos H, Kinnunen M, Salokas K, Tan Z, Liu X, Yadav L, Zhang Q, Wei 
            GH, Varjosalo M
  TITLE     Human transcription factor protein interaction networks.
  JOURNAL   Nat Commun. 2022 Feb 9;13(1):766. doi: 10.1038/s41467-022-28341-5.
  PUBMED    35140242
REFERENCE   2  (bases 1 to 7563)
  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 
            Commun."; date: "2022-02-9"; volume: "13(1):766. doi"; pages: " 
            10.1038/s41467-022-28341-5"
COMMENT     Sequence Label: BirA-Myc_N_ETS1
FEATURES             Location/Qualifiers
     source          1..7563
                     /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"
     CDS             830..859
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(863..1210,1211..1213,1214..1822)
                     /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"
     protein_bind    1865..1889
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3219..3243)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3281..3298)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3287..3511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3644..3663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3795..3842
                     /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             3850..4872
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3850..4872
                     /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    5002..5123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5039..5058)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5093..5112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5172..5188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5172..5188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5185..5207)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5196..5212
                     /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(5220..5226,5227..5244,5245..5250))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5265..5286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5403..5420)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5574..6162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5654..5673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6333..7124,7125..7193))
                     /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     6956..6975
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7194..7298)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7373..7392)
                     /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 cagcacagtg gcggccgctc gagaccatgg agcagaagct
      841 gatcagcgaa gaggacctgg acaaggacaa caccgtgccc ctgaagctga tcgccctgct
      901 ggccaacggc gagttccact ctggcgagca gctgggcgag acactgggca tgagcagagc
      961 cgccatcaac aagcacatcc agaccctgcg ggactggggc gtggacgtgt tcacagtgcc
     1021 cggcaagggc tacagcctgc ccgagcccat ccagctgctg aacgccaagc agatcctggg
     1081 ccagctggat ggcggatctg tggccgtgct gcccgtgatc gacagcacca accagtatct
     1141 gctcgaccgg atcggcgagc tgaagtccgg cgacgcctgt atcgccgagt accagcaggc
     1201 cggcagaggc ggcaggggca gaaagtggtt cagccccttc ggcgccaacc tgtacctgag
     1261 catgttctgg agactggaac agggcccagc cgccgctatt ggcctgagcc tggtcatcgg
     1321 catcgtgatg gccgaggtgc tgcggaagct gggcgccgac aaagtgcgcg tgaagtggcc
     1381 caacgacctg tatctgcagg accggaagct ggccggcatc ctggtggaac tgaccggcaa
     1441 aacaggcgac gccgcccaga tcgtgatcgg cgctggcatc aacatggcca tgcggcgggt
     1501 ggaagagagc gtggtcaacc agggctggat caccctgcag gaagccggca tcaacctgga
     1561 ccggaacacc ctggccgcca tgctgatcag agagctgaga gccgccctgg aactgttcga
     1621 gcaggaaggc ctggccccct acctgagcag atgggagaag ctggacaact tcatcaaccg
     1681 gcccgtgaaa ctgatcatcg gcgacaaaga gatcttcggc atcagccggg gcatcgacaa
     1741 gcagggggcc ctgctcctgg aacaggacgg catcatcaag ccctggatgg gcggcgagat
     1801 cagcctgcgg agcgccgaga agggcggagg ctctggcggc ggaagcggcg gaggcagcga
     1861 tatcacaagt ttgtacaaaa aagcaggctc caccatgaag gcggccgtcg atctcaagcc
     1921 gactctcacc atcatcaaga cggaaaaagt cgatctggag cttttcccct ccccggatat
     1981 ggaatgtgca gatgtcccac tattaactcc aagcagcaaa gaaatgatgt ctcaagcatt
     2041 aaaagctact ttcagtggtt tcactaaaga acagcaacga ctggggatcc caaaagaccc
     2101 ccggcagtgg acagaaaccc atgttcggga ctgggtgatg tgggctgtga atgaattcag
     2161 cctgaaaggt gtagacttcc agaagttctg tatgaatgga gcagccctct gcgccctggg
     2221 taaagactgc tttctcgagc tggccccaga ctttgttggg gacatcttat gggaacatct
     2281 agagatcctg cagaaagagg atgtgaaacc atatcaagtt aatggagtca acccagccta
     2341 tccagaatcc cgctatacct cggattactt cattagctat ggtattgagc atgcccagtg
     2401 tgttccacca tcggagttct cagagcccag cttcatcaca gagtcctatc agacgctcca
     2461 tcccatcagc tcggaagagc tcctctccct caagtatgag aatgactacc cctcggtcat
     2521 tctccgagac cctctccaga cagacacctt gcagaatgac tactttgcta tcaaacaaga
     2581 agtcgtcacc ccagacaaca tgtgcatggg gaggaccagt cgtggtaaac tcgggggcca
     2641 ggactctttt gaaagcatag agagctacga tagttgtgat cgcctcaccc agtcctggag
     2701 cagccagtca tctttcaaca gcctgcagcg tgttccctcc tatgacagct tcgactcaga
     2761 ggactatccg gctgccctgc ccaaccacaa gcccaagggc accttcaagg actatgtgcg
     2821 ggaccgtgct gacctcaata aggacaagcc tgtcattcct gctgctgccc tagctggcta
     2881 cacaggcagt ggaccaatcc agctatggca gtttcttctg gaattactca ctgataaatc
     2941 ctgtcagtct tttatcagct ggacaggaga tggctgggaa ttcaaacttt ctgacccaga
     3001 tgaggtggcc aggagatggg gaaagaggaa aaacaaacct aagatgaatt atgagaaact
     3061 gagccgtggc ctacgctact attacgacaa aaacatcatc cacaagacag cggggaaacg
     3121 ctacgtgtac cgctttgtgt gtgacctgca gagcctgctg gggtacaccc ctgaggagct
     3181 gcacgccatg ctggacgtca agccagatgc cgacgaggac ccagctttct tgtacaaagt
     3241 ggtgacgtaa gctaggggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3301 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3361 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3421 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3481 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3541 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3601 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3661 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3721 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3781 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact
     3841 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     3901 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     3961 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     4021 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     4081 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     4141 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     4201 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     4261 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     4321 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     4381 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     4441 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     4501 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     4561 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     4621 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     4681 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     4741 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     4801 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     4861 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4921 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4981 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5041 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5101 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5161 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5221 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5281 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5341 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5401 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5461 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5521 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5581 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5641 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5701 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5761 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5821 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5881 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5941 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6001 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6061 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6121 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6181 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6241 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6301 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6361 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6421 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6481 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6541 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6601 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6661 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6721 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6781 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6841 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6901 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6961 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7021 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7081 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7141 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7201 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7261 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7321 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     7381 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7441 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7501 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     7561 gtt
//
