LOCUS       sequence-370417-        7596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human NFIC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7596)
  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 7596)
  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_NFIC
FEATURES             Location/Qualifiers
     source          1..7596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        443..646
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     597..617
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    648..666
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    669..687
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     697..721
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(889..1239,1240..1242,1243..1851)
                     /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             1852..1878
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1909..1932
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1969..1992
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    2005..2029
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3314..3338)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3376..3393)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3382..3606
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3739..3758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3890..3937
                     /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             3945..4967
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3945..4967
                     /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    5097..5218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5134..5153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5188..5207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5267..5283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5267..5283)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5280..5302)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5291..5307
                     /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(5315..5321,5322..5339,5340..5345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5360..5381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5498..5515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5669..6257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5749..5768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6428..7219,7220..7288))
                     /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     7051..7070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7289..7393)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7468..7487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
       61 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      121 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      181 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      241 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      301 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      361 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      421 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      481 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      541 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      601 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
      661 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
      721 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
      781 cctccggact ctagcgttta aacttaagct tggtaccgag ctcggatcca ctagtccagt
      841 gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagaccat gaaggacaac
      901 accgtgcccc tgaagctgat cgccctgctg gccaacggcg agttccactc tggcgagcag
      961 ctgggcgaga cactgggcat gagcagagcc gccatcaaca agcacatcca gaccctgcgg
     1021 gactggggcg tggacgtgtt cacagtgccc ggcaagggct acagcctgcc cgagcccatc
     1081 cagctgctga acgccaagca gatcctgggc cagctggatg gcggatctgt ggccgtgctg
     1141 cccgtgatcg acagcaccaa ccagtatctg ctcgaccgga tcggcgagct gaagtccggc
     1201 gacgcctgta tcgccgagta ccagcaggcc ggcagaggcg gcaggggcag aaagtggttc
     1261 agccccttcg gcgccaacct gtacctgagc atgttctgga gactggaaca gggcccagcc
     1321 gccgctattg gcctgagcct ggtcatcggc atcgtgatgg ccgaggtgct gcggaagctg
     1381 ggcgccgaca aagtgcgcgt gaagtggccc aacgacctgt atctgcagga ccggaagctg
     1441 gccggcatcc tggtggaact gaccggcaaa acaggcgacg ccgcccagat cgtgatcggc
     1501 gctggcatca acatggccat gcggcgggtg gaagagagcg tggtcaacca gggctggatc
     1561 accctgcagg aagccggcat caacctggac cggaacaccc tggccgccat gctgatcaga
     1621 gagctgagag ccgccctgga actgttcgag caggaaggcc tggcccccta cctgagcaga
     1681 tgggagaagc tggacaactt catcaaccgg cccgtgaaac tgatcatcgg cgacaaagag
     1741 atcttcggca tcagccgggg catcgacaag cagggggccc tgctcctgga acaggacggc
     1801 atcatcaagc cctggatggg cggcgagatc agcctgcgga gcgccgagaa gtacccatac
     1861 gatgttcctg actatgccgg taccgagctc ggatccacca tggctagctg gagccacccg
     1921 cagttcgaga aaggtggagg ttccggaggt ggatcgggag gtggatcgtg gagccacccg
     1981 cagttcgaaa aagcggccga tatcacaagt ttgtacaaaa aagttggcat gtattcgtcc
     2041 ccgctctgcc tcacccagga tgagttccac ccgttcatcg aggccctgct gcctcacgtc
     2101 cgcgccttcg cctacacctg gttcaacctg caggcgcgga agcgcaagta cttcaagaag
     2161 cacgagaagc ggatgtcgaa ggacgaggag cgtgcggtca aggacgagct gctgggcgag
     2221 aagcccgagg tcaagcagaa gtgggcgtcg cggctgctgg ccaagctgcg caaggacatc
     2281 cggcccgagt gccgcgagga cttcgtgctg agcatcaccg gcaagaaggc gccgggctgc
     2341 gtgctctcca accccgacca gaagggcaag atgcggcgca tcgactgtct ccggcaggcg
     2401 gacaaggtgt ggcggctgga cctggtcatg gtcatcctgt tcaagggcat cccgctggag
     2461 agcaccgacg gcgagcgcct ggtcaaggct gcgcagtgcg gtcacccggt cctgtgcgtg
     2521 cagccgcacc acattggcgt ggccgtcaag gagctggacc tctacctggc ctacttcgtg
     2581 cgtgagcgag atgcagagca aagcggcagt ccccggacag ggatgggctc tgaccaggag
     2641 gacagcaagc ccatcacgct ggacacgacc gacttccagg agagctttgt cacctccggc
     2701 gtgttcagcg tcactgagct catccaagtg tcccggacac ccgtggtgac tggaacagga
     2761 cccaacttct ccctggggga gctgcagggg cacctggcat acgacctgaa cccagccagc
     2821 actggcctca gaagaacgct gcccagcacc tcctccagtg ggagcaagcg gcacaaatcg
     2881 ggctcgatgg aggaagacgt ggacacgagc cctggcggcg attactacac ttcgcccagc
     2941 tcgcccacga gtagcagccg caactggacg gaggacatgg aaggaggcat ctcgtccccg
     3001 gtgaagaaga cagagatgga caagtcacca ttcaacagcc cgtcccccca ggactctccc
     3061 cgcctctcca gcttcaccca gcaccaccgg cccgtcatcg ccgtgcacag cgggatcgcc
     3121 cggagcccac acccgtcctc cgctctgcat ttccctacga cgtccatcct accccagacg
     3181 gcctccacct acttccccca cacggccatc cgctacccac ctcatctcaa cccccaggac
     3241 ccgctcaaag atcttgtctc gctggcctgc gacccagcca gccagcaacc tggaccgtcc
     3301 tggtatctgg gatacccaac tttcttgtac aaagtggtga cgtaagctag gggcccgttt
     3361 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3421 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3481 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3541 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3601 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3661 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3721 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3781 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3841 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat
     3901 tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca
     3961 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
     4021 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
     4081 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
     4141 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
     4201 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
     4261 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
     4321 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
     4381 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
     4441 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
     4501 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
     4561 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
     4621 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
     4681 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
     4741 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
     4801 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
     4861 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
     4921 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag
     4981 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5041 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5101 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5161 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5221 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5281 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5341 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5401 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5461 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5521 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5581 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5641 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5701 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5761 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5821 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5881 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5941 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6001 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6061 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6121 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6181 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6241 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6301 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6361 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6421 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6481 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6541 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6601 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6661 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6721 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6781 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6841 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6901 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6961 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7021 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7081 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7141 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7201 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7261 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7321 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7381 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7441 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7501 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     7561 tacaacaagg caaggcttga ccgacaattg catgaa
//
