LOCUS       sequence-370461-        7380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human LHX8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  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 7380)
  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_LHX8
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        89..468
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        469..672
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     623..643
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    674..692
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    695..713
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     723..747
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(915..1265,1266..1268,1269..1877)
                     /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             1878..1904
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1935..1958
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1995..2018
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    2031..2055
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3124..3148)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3186..3203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3192..3416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3549..3568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3700..3747
                     /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             3755..4777
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3755..4777
                     /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    4907..5028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4944..4963)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4998..5017
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5077..5093)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5077..5093)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5090..5112)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5101..5117
                     /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(5125..5131,5132..5149,5150..5155))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5170..5191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5308..5325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5479..6067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5559..5578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6238..7029,7030..7098))
                     /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     6861..6880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7099..7203)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7278..7297)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
       61 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      121 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      181 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      241 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      301 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      361 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      421 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      481 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      541 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      601 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      661 ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga
      721 cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      781 catagaagac accgggaccg atccagcctc cggactctag cgtttaaact taagcttggt
      841 accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca gcacagtggc
      901 ggccgctcga gaccatgaag gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca
      961 acggcgagtt ccactctggc gagcagctgg gcgagacact gggcatgagc agagccgcca
     1021 tcaacaagca catccagacc ctgcgggact ggggcgtgga cgtgttcaca gtgcccggca
     1081 agggctacag cctgcccgag cccatccagc tgctgaacgc caagcagatc ctgggccagc
     1141 tggatggcgg atctgtggcc gtgctgcccg tgatcgacag caccaaccag tatctgctcg
     1201 accggatcgg cgagctgaag tccggcgacg cctgtatcgc cgagtaccag caggccggca
     1261 gaggcggcag gggcagaaag tggttcagcc ccttcggcgc caacctgtac ctgagcatgt
     1321 tctggagact ggaacagggc ccagccgccg ctattggcct gagcctggtc atcggcatcg
     1381 tgatggccga ggtgctgcgg aagctgggcg ccgacaaagt gcgcgtgaag tggcccaacg
     1441 acctgtatct gcaggaccgg aagctggccg gcatcctggt ggaactgacc ggcaaaacag
     1501 gcgacgccgc ccagatcgtg atcggcgctg gcatcaacat ggccatgcgg cgggtggaag
     1561 agagcgtggt caaccagggc tggatcaccc tgcaggaagc cggcatcaac ctggaccgga
     1621 acaccctggc cgccatgctg atcagagagc tgagagccgc cctggaactg ttcgagcagg
     1681 aaggcctggc cccctacctg agcagatggg agaagctgga caacttcatc aaccggcccg
     1741 tgaaactgat catcggcgac aaagagatct tcggcatcag ccggggcatc gacaagcagg
     1801 gggccctgct cctggaacag gacggcatca tcaagccctg gatgggcggc gagatcagcc
     1861 tgcggagcgc cgagaagtac ccatacgatg ttcctgacta tgccggtacc gagctcggat
     1921 ccaccatggc tagctggagc cacccgcagt tcgagaaagg tggaggttcc ggaggtggat
     1981 cgggaggtgg atcgtggagc cacccgcagt tcgaaaaagc ggccgatatc acaagtttgt
     2041 acaaaaaagt tggcatgcag attctgagca ggtgtcaggg gctcatgtca gaggagtgcg
     2101 ggcggactac agccctggcg gccgggagga ctcgcaaagg cgccggggaa gagggactgg
     2161 tgagccccga gggagcgggg gacgaggact cgtgctcctc ctcggccccg ctgtccccgt
     2221 cgtcctcgcc ccggtccatg gcctcgggct ccggctgccc tcctggcaag tgtgtgtgca
     2281 acagttgcgg cctggagatc gtggacaaat accttctcaa ggtgaatgac ctatgctggc
     2341 atgtccggtg tctctcctgc agtgtttgca gaacctccct aggaaggcac accagctgtt
     2401 atattaaaga caaagacatt ttctgcaaac ttgattattt cagaaggtat ggaactcgct
     2461 gctctcgatg tgggagacac atccattcta ctgactgggt ccggagagcc aaggggaatg
     2521 tctatcactt ggcatgcttt gcctgctttt cctgcaaaag gcaactttcc acaggagagg
     2581 agtttgcttt ggtggaagag aaagtcctct gcagagtaca ttatgactgc atgctggata
     2641 atttaaaaag agaagtagaa aatgggaatg ggattagtgt ggaaggtgcc ctcctcacag
     2701 agcaagatgt taaccatcca aaaccagcaa aaagagctcg gaccagcttt acagcagatc
     2761 agcttcaggt tatgcaagca caatttgctc aggacaacaa cccagatgca cagacactcc
     2821 agaaattggc agaaaggaca ggcttgagca gacgtgtgat acaggtgtgg tttcagaatt
     2881 gtagagcacg ccacaagaaa cacgtcagtc ctaatcactc atcctccacc ccagtcacag
     2941 cagccccacc ctccaggctg tctccaccca tgttagaaga aatggcttat tctgcctacg
     3001 tgccccaaga tggaacgatg ttaactgcgc tgcatagtta tatggatgct cattcaccaa
     3061 caactcttgg actccagccc ttgttacccc attcaatgac acaactgcca ataagtcata
     3121 cctacccaac tttcttgtac aaagtggtga cgtaagctag gggcccgttt aaacccgctg
     3181 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3241 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3301 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3361 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3421 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3481 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3541 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3601 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3661 ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc
     3721 ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc
     3781 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3841 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3901 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     3961 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg
     4021 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     4081 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     4141 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     4201 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     4261 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4321 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4381 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4441 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4501 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     4561 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4621 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4681 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4741 ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc
     4801 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4861 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4921 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4981 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5041 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5101 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5161 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5221 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5281 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5341 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5401 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5641 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6121 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6181 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6241 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6301 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6361 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6421 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6481 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6541 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6601 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6661 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6721 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6781 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6841 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6901 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6961 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7021 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7081 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7141 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7201 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7261 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7321 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
//
