LOCUS       sequence-370414-        7548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human LHX3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7548)
  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 7548)
  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_LHX3
FEATURES             Location/Qualifiers
     source          1..7548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             54..80
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             111..134
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             171..194
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    207..231
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1468..1492)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1530..1547)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1536..1760
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1893..1912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2044..2091
                     /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             2099..3121
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2099..3121
                     /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    3251..3372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3288..3307)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3342..3361
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3421..3437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3421..3437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3434..3456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3445..3461
                     /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(3469..3475,3476..3493,3494..3499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3514..3535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3652..3669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3823..4411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3903..3922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4582..5373,5374..5442))
                     /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     5205..5224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5443..5547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5622..5641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5813..6192
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6193..6396
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6347..6367
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6398..6416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6419..6437
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6447..6471
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(6639..6989,6990..6992,6993..7548,1..53)
                     /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"
ORIGIN
        1 gcatcatcaa gccctggatg ggcggcgaga tcagcctgcg gagcgccgag aagtacccat
       61 acgatgttcc tgactatgcc ggtaccgagc tcggatccac catggctagc tggagccacc
      121 cgcagttcga gaaaggtgga ggttccggag gtggatcggg aggtggatcg tggagccacc
      181 cgcagttcga aaaagcggcc gatatcacaa gtttgtacaa aaaagcagaa gggccgtcaa
      241 ggcccaccat ggaggcgcgc ggggagctgg gcccggcccg ggagtcggcg ggaggcgacc
      301 tgctgctagc actgctggcg cggagggcgg acctgcgccg agagatcccg ctgtgcgctg
      361 gctgtgacca gcacatcctg gaccgcttca tcctcaaggc tctggaccgc cactggcaca
      421 gcaagtgtct caagtgcagc gactgccaca cgccactggc cgagcgctgc ttcagccgag
      481 gggagagcgt ttactgcaag gacgactttt tcaagcgctt cgggaccaag tgcgccgcgt
      541 gccagctggg catcccgccc acgcaggtgg tgcgccgcgc ccaggacttc gtgtaccacc
      601 tgcactgctt tgcctgcgtc gtgtgcaagc ggcagctggc cacgggcgac gagttctacc
      661 tcatggagga cagccggctc gtgtgcaagg cggactacga aaccgccaag cagcgagagg
      721 ccgaggccac ggccaagcgg ccgcgcacga ccatcaccgc caagcagctg gagacgctga
      781 agagcgctta caacacctcg cccaagccgg cgcgccacgt gcgcgagcag ctctcgtccg
      841 agacgggcct ggacatgcgc gtggtgcagg tttggttcca gaaccgccgg gccaaggaga
      901 agaggctgaa gaaggacgcc ggccggcagc gctgggggca gtatttccgc aacatgaagc
      961 gctcccgcgg cggctccaag tcggacaagg acagcgttca ggaggggcag gacagcgacg
     1021 ctgaggtctc cttccccgat gagccttcct tggcggaaat gggcccggcc aatggcctct
     1081 acgggagctt gggggaaccc acccaggcct tgggccggcc ctcgggagcc ctgggcaact
     1141 tctccctgga gcatggaggc ctggcaggcc cagagcagta ccgagagctg cgtcccggca
     1201 gcccctacgg tgtcccccca tcccccgccg ccccgcagag cctccctggc ccccagcccc
     1261 tcctctccag cctggtgtac ccagacacca gcttgggcct tgtgccctcg ggagcccccg
     1321 gcgggccccc acccatgagg gtgctggcag ggaacggacc cagttctgac ctatccacgg
     1381 ggagcagcgg gggttacccc gacttccctg ccagccccgc ctcctggctg gatgaggtag
     1441 accacgctca gttctcaggc ctcatgggcc cagctttctt gtacaaagtg gtgacgtaag
     1501 ctaggggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     1561 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1621 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1681 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1741 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     1801 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1861 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1921 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1981 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     2041 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     2101 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     2161 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     2221 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     2281 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     2341 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     2401 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     2461 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     2521 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     2581 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     2641 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     2701 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     2761 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     2821 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     2881 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     2941 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     3001 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     3061 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     3121 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3181 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3241 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3301 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3361 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     3421 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3481 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3541 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3601 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3661 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3721 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3781 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3841 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3901 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3961 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4021 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4081 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4141 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4201 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4261 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4321 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4381 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4441 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4501 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4561 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4621 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4681 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4741 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4801 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4861 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4921 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4981 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5041 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5101 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5161 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5221 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5281 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5341 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5401 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5461 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5521 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     5581 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     5641 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     5701 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     5761 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     5821 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5881 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5941 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6001 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6061 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6121 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6181 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6241 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6301 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6361 aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga tagagatctc
     6421 cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat cgcctggaga
     6481 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
     6541 ctagcgttta aacttaagct tggtaccgag ctcggatcca ctagtccagt gtggtggaat
     6601 tctgcagata tccagcacag tggcggccgc tcgagaccat gaaggacaac accgtgcccc
     6661 tgaagctgat cgccctgctg gccaacggcg agttccactc tggcgagcag ctgggcgaga
     6721 cactgggcat gagcagagcc gccatcaaca agcacatcca gaccctgcgg gactggggcg
     6781 tggacgtgtt cacagtgccc ggcaagggct acagcctgcc cgagcccatc cagctgctga
     6841 acgccaagca gatcctgggc cagctggatg gcggatctgt ggccgtgctg cccgtgatcg
     6901 acagcaccaa ccagtatctg ctcgaccgga tcggcgagct gaagtccggc gacgcctgta
     6961 tcgccgagta ccagcaggcc ggcagaggcg gcaggggcag aaagtggttc agccccttcg
     7021 gcgccaacct gtacctgagc atgttctgga gactggaaca gggcccagcc gccgctattg
     7081 gcctgagcct ggtcatcggc atcgtgatgg ccgaggtgct gcggaagctg ggcgccgaca
     7141 aagtgcgcgt gaagtggccc aacgacctgt atctgcagga ccggaagctg gccggcatcc
     7201 tggtggaact gaccggcaaa acaggcgacg ccgcccagat cgtgatcggc gctggcatca
     7261 acatggccat gcggcgggtg gaagagagcg tggtcaacca gggctggatc accctgcagg
     7321 aagccggcat caacctggac cggaacaccc tggccgccat gctgatcaga gagctgagag
     7381 ccgccctgga actgttcgag caggaaggcc tggcccccta cctgagcaga tgggagaagc
     7441 tggacaactt catcaaccgg cccgtgaaac tgatcatcgg cgacaaagag atcttcggca
     7501 tcagccgggg catcgacaag cagggggccc tgctcctgga acaggacg
//
