LOCUS       sequence-333406-        7416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HME1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7416)
  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 7416)
  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_HME1
FEATURES             Location/Qualifiers
     source          1..7416
                     /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(3072..3096)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3134..3151)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3140..3364
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3497..3516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3648..3695
                     /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             3703..4725
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3703..4725
                     /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    4855..4976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4892..4911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4946..4965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5025..5041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5025..5041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5038..5060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5049..5065
                     /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(5073..5079,5080..5097,5098..5103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5118..5139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5256..5273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5427..6015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5507..5526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6186..6977,6978..7046))
                     /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     6809..6828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7047..7151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7226..7245)
                     /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 caccatggaa gaacagcagc cggaacctaa
     1921 aagtcagcgc gactcggccc tcggcgcggc ggcggcggcg actccgggcg gcctcagcct
     1981 gagcctcagt ccgggcgcca gcggcagcag cggcagcggc agcgatggag acagcgtgcc
     2041 ggtgtccccg cagcctgcgc ccccctcgcc gcccgcggcg ccttgcctgc cgcccctggc
     2101 ccaccacccg cacctccccc cacacccccc gcccccgccg cctcagcatc tcgcggcgcc
     2161 tgctcaccag ccgcagccag cggcccagct gcaccgcacc accaactttt tcatcgacaa
     2221 catcctgagg ccggacttcg gctgcaaaaa ggagcagccg ccaccgcagc ttctggtggc
     2281 tgcggcggcc agaggaggcg caggaggagg aggccgggtc gagcgtgaca gaggccagac
     2341 tgccgcaggt agagaccctg tccacccgtt gggcacccgg gcgccaggcg ctgcctcgct
     2401 cctgtgcgcc ccggacgcga actgtggccc acccgacggc tcccagccag ccgccgccgg
     2461 cgcgggcgcg tctaaagctg ggaacccggc tgcggcggcg gcggcggccg cggcggcagt
     2521 ggcggcggcg gcggcggccg cagcagccaa gccctcggac accggtggcg gcggcagtgg
     2581 aggcggcgcg gggagccccg gagcgcaggg caccaaatac ccggagcacg gcaacccggc
     2641 tatcctactt atgggctcag ccaacggcgg gcccgtggtc aaaactgact cgcagcagcc
     2701 tctcgtatgg cccgcctggg tgtactgcac acgttattcg gatcgtccat cctccggtcc
     2761 gcgcaccagg aagctgaaga agaagaagaa cgagaaggag gacaagcggc cgcggaccgc
     2821 gttcacggcc gagcagctgc agagactcaa ggcggagttc caggcaaacc gctacatcac
     2881 ggagcagcgg cggcagaccc tggcccagga actcagcctc aacgagtccc agatcaagat
     2941 ctggttccag aacaagcgcg ccaagatcaa gaaagccaca ggcatcaaga acggcctggc
     3001 gctgcacctc atggcccagg gactgtacaa ccactccacc accacggtcc aggacaaaga
     3061 cgagagcgag gacccagctt tcttgtacaa agtggtgacg taagctaggg gcccgtttaa
     3121 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3181 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3241 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3301 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3361 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3421 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3481 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3541 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3601 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc
     3661 cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc tgaactcacc
     3721 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     3781 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     3841 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     3901 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
     3961 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     4021 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
     4081 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     4141 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     4201 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     4261 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     4321 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     4381 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     4441 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     4501 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     4561 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     4621 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     4681 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt actacgagat
     4741 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4801 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4861 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4921 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4981 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5041 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5101 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5161 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5221 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5281 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5341 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5401 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5461 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5521 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5581 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5641 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5701 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5761 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5821 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5881 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5941 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6001 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6061 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6121 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6181 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6241 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6301 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6361 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6421 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6481 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6541 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6601 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6661 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6721 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6781 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6841 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6901 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6961 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7021 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7081 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7141 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7201 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7261 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7321 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     7381 cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
