LOCUS       sequence-370378-        7050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human ISX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7050)
  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 7050)
  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_ISX
FEATURES             Location/Qualifiers
     source          1..7050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..33)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(187..775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(267..286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(946..1737,1738..1806))
                     /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     1569..1588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1807..1911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1986..2005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2177..2556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2557..2760
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2711..2731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2762..2780
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2783..2801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2811..2835
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3003..3353,3354..3356,3357..3965)
                     /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             3966..3992
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4023..4046
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4083..4106
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    4119..4143
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4882..4906)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4944..4961)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4950..5174
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5307..5326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5458..5505
                     /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             5513..6535
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5513..6535
                     /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    6665..6786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6702..6721)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6756..6775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6835..6851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6835..6851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6848..6870)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6859..6875
                     /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(6883..6889,6890..6907,6908..6913))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6928..6949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
       61 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1921 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     1981 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     2041 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     2101 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     2161 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     2221 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2281 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2341 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2401 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2461 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2521 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2581 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2641 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     2701 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     2761 ctccctatca gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta
     2821 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     2881 cgggaccgat ccagcctccg gactctagcg tttaaactta agcttggtac cgagctcgga
     2941 tccactagtc cagtgtggtg gaattctgca gatatccagc acagtggcgg ccgctcgaga
     3001 ccatgaagga caacaccgtg cccctgaagc tgatcgccct gctggccaac ggcgagttcc
     3061 actctggcga gcagctgggc gagacactgg gcatgagcag agccgccatc aacaagcaca
     3121 tccagaccct gcgggactgg ggcgtggacg tgttcacagt gcccggcaag ggctacagcc
     3181 tgcccgagcc catccagctg ctgaacgcca agcagatcct gggccagctg gatggcggat
     3241 ctgtggccgt gctgcccgtg atcgacagca ccaaccagta tctgctcgac cggatcggcg
     3301 agctgaagtc cggcgacgcc tgtatcgccg agtaccagca ggccggcaga ggcggcaggg
     3361 gcagaaagtg gttcagcccc ttcggcgcca acctgtacct gagcatgttc tggagactgg
     3421 aacagggccc agccgccgct attggcctga gcctggtcat cggcatcgtg atggccgagg
     3481 tgctgcggaa gctgggcgcc gacaaagtgc gcgtgaagtg gcccaacgac ctgtatctgc
     3541 aggaccggaa gctggccggc atcctggtgg aactgaccgg caaaacaggc gacgccgccc
     3601 agatcgtgat cggcgctggc atcaacatgg ccatgcggcg ggtggaagag agcgtggtca
     3661 accagggctg gatcaccctg caggaagccg gcatcaacct ggaccggaac accctggccg
     3721 ccatgctgat cagagagctg agagccgccc tggaactgtt cgagcaggaa ggcctggccc
     3781 cctacctgag cagatgggag aagctggaca acttcatcaa ccggcccgtg aaactgatca
     3841 tcggcgacaa agagatcttc ggcatcagcc ggggcatcga caagcagggg gccctgctcc
     3901 tggaacagga cggcatcatc aagccctgga tgggcggcga gatcagcctg cggagcgccg
     3961 agaagtaccc atacgatgtt cctgactatg ccggtaccga gctcggatcc accatggcta
     4021 gctggagcca cccgcagttc gagaaaggtg gaggttccgg aggtggatcg ggaggtggat
     4081 cgtggagcca cccgcagttc gaaaaagcgg ccgatatcac aagtttgtac aaaaaagttg
     4141 gcaccatgtg tgctgaggtg ggccctgctc tctgcagggg tatggagaga aatagcttgg
     4201 ggtgctgtga ggccccgaag aagctgggcc tgtccttctc catcgaggcg atcctaaaga
     4261 ggcctgccag gaggagtgat atggacagac cagaagggcc aggtgaagag ggctccggag
     4321 aagctgcggc ctcaggctct gggctagaaa agcctccaaa ggaccagccc caggaaggaa
     4381 ggaagagcaa gcggagggtt cgtaccacct tcaccactga gcagctgcat gagctggaga
     4441 agatcttcca ctttacccac tacccagacg ttcacatccg cagccagctg gcagccagga
     4501 tcaacctccc agaagctcgg gtgcagatct ggttccagaa tcagcgagcc aagtggcgga
     4561 agcaggagaa gattggcaac ctgggggctc cacagcagct gagtgaagcc agtgtggccc
     4621 tgcccacaaa tctggatgtg gctgggccca cgtggacatc cactgctctg cgcaggctgg
     4681 ctcctcccac gagctgttgt ccatcggctc aagatcagct ggcctctgcc tggttccctg
     4741 cctggatcac cctcctccca gcgcacccat gggaaacaca gcctgtccca ggtcttccca
     4801 tccatcaaac ttgcatccct gtgctatgca tccttccacc tccacacccc aaatggggca
     4861 gcatctgtgc tacttcaaca ttgccaactt tcttgtacaa agtggtgacg taagctaggg
     4921 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4981 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5041 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5101 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5161 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     5221 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5281 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5341 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5401 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa
     5461 gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc
     5521 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     5581 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     5641 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     5701 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     5761 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     5821 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     5881 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     5941 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     6001 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     6061 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     6121 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     6181 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     6241 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     6301 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     6361 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     6421 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     6481 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     6541 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6601 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6661 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6781 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6841 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6901 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7021 acgcgcgggg agaggcggtt tgcgtattgg
//
