LOCUS       sequence-370481-        7077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human ELF5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7077)
  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 7077)
  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: MAC_N_ELF5
FEATURES             Location/Qualifiers
     source          1..7077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        134..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        514..717
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     668..688
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    719..737
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    740..758
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     768..792
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(960..1310,1311..1313,1314..1922)
                     /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="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1923..1949
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1980..2003
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2040..2063
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    2076..2100
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2866..2890)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2928..2945)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2934..3158
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3291..3310)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3442..3489
                     /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             3497..4519
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3497..4519
                     /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    4649..4770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4686..4705)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4740..4759
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4819..4835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4819..4835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4832..4854)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4843..4859
                     /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(4867..4873,4874..4891,4892..4897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4912..4933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5050..5067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5221..5809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5301..5320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5980..6771,6772..6840))
                     /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     6603..6622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6841..6945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7020..7039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
       61 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      121 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      541 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      601 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      661 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      721 cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg
      781 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      841 gaccgatcca gcctccggac tctagcgttt aaacttaagc ttggtaccga gctcggatcc
      901 actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagacca
      961 tgaaggacaa caccgtgccc ctgaagctga tcgccctgct ggccaacggc gagttccact
     1021 ctggcgagca gctgggcgag acactgggca tgagcagagc cgccatcaac aagcacatcc
     1081 agaccctgcg ggactggggc gtggacgtgt tcacagtgcc cggcaagggc tacagcctgc
     1141 ccgagcccat ccagctgctg aacgccaagc agatcctggg ccagctggat ggcggatctg
     1201 tggccgtgct gcccgtgatc gacagcacca accagtatct gctcgaccgg atcggcgagc
     1261 tgaagtccgg cgacgcctgt atcgccgagt accagcaggc cggcagaggc ggcaggggca
     1321 gaaagtggtt cagccccttc ggcgccaacc tgtacctgag catgttctgg agactggaac
     1381 agggcccagc cgccgctatt ggcctgagcc tggtcatcgg catcgtgatg gccgaggtgc
     1441 tgcggaagct gggcgccgac aaagtgcgcg tgaagtggcc caacgacctg tatctgcagg
     1501 accggaagct ggccggcatc ctggtggaac tgaccggcaa aacaggcgac gccgcccaga
     1561 tcgtgatcgg cgctggcatc aacatggcca tgcggcgggt ggaagagagc gtggtcaacc
     1621 agggctggat caccctgcag gaagccggca tcaacctgga ccggaacacc ctggccgcca
     1681 tgctgatcag agagctgaga gccgccctgg aactgttcga gcaggaaggc ctggccccct
     1741 acctgagcag atgggagaag ctggacaact tcatcaaccg gcccgtgaaa ctgatcatcg
     1801 gcgacaaaga gatcttcggc atcagccggg gcatcgacaa gcagggggcc ctgctcctgg
     1861 aacaggacgg catcatcaag ccctggatgg gcggcgagat cagcctgcgg agcgccgaga
     1921 agtacccata cgatgttcct gactatgccg gtaccgagct cggatccacc atggctagct
     1981 ggagccaccc gcagttcgag aaaggtggag gttccggagg tggatcggga ggtggatcgt
     2041 ggagccaccc gcagttcgaa aaagcggccg atatcacaag tttgtacaaa aaagttggca
     2101 tgttggactc ggtgacacac agcaccttcc tgcctaatgc atccttctgc gatcccctga
     2161 tgtcgtggac tgatctgttc agcaatgaag agtactaccc tgcctttgag catcagacag
     2221 cctgtgactc atactggaca tcagtccacc ctgaatactg gactaagcgc catgtgtggg
     2281 agtggctcca gttctgctgc gaccagtaca agttggacac caattgcatc tccttctgca
     2341 acttcaacat cagtggcctg cagctgtgca gcatgacaca ggaggagttc gtcgaggcag
     2401 ctggcctctg cggcgagtac ctgtacttca tcctccagaa catccgcaca caaggttact
     2461 ccttttttaa tgacgctgaa gaaagcaagg ccaccatcaa agactatgct gattccaact
     2521 gcttgaaaac aagtggcatc aaaagtcaag actgtcacag tcatagtaga acaagcctcc
     2581 aaagttctca tctatgggaa tttgtacgag acctgcttct atctcctgaa gaaaactgtg
     2641 gcattctgga atgggaagat agggaacaag gaatttttcg ggtggttaaa tcggaagccc
     2701 tggcaaagat gtggggacaa aggaagaaaa atgacagaat gacatatgaa aagttgagca
     2761 gagccctgag atactactat aaaacaggaa ttttggagcg ggttgaccga aggttagtgt
     2821 acaaatttgg aaaaaatgca cacgggtggc aggaagacaa gctatgccca actttcttgt
     2881 acaaagtggt gacgtaagct aggggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     2941 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3001 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3061 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3121 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3181 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3241 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3301 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3361 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3421 gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat
     3481 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3541 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3601 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     3661 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     3721 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     3781 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     3841 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     3901 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     3961 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4021 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4081 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4141 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4201 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4261 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4321 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4381 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4441 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4501 tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga
     4561 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4621 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4681 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4741 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4801 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4861 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4921 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4981 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5281 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5341 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5401 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5461 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5521 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5581 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5641 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5701 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5761 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5821 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5881 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5941 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6001 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6061 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6421 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6961 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7021 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttg
//
