LOCUS       sequence-333749-        7230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TLX1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7230)
  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 7230)
  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_TLX1
FEATURES             Location/Qualifiers
     source          1..7230
                     /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(2886..2910)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2948..2965)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2954..3178
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3311..3330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3462..3509
                     /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             3517..4539
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3517..4539
                     /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    4669..4790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4706..4725)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4760..4779
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4839..4855)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4839..4855)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4852..4874)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4863..4879
                     /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(4887..4893,4894..4911,4912..4917))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4932..4953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5070..5087)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5241..5829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5321..5340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6000..6791,6792..6860))
                     /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     6623..6642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6861..6965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7040..7059)
                     /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 caccatggag cacctgggtc cgcaccacct
     1921 ccacccgggt cacgcagagc ccattagctt cggcatcgac cagatcctca acagcccgga
     1981 ccagggtggc tgcatgggac ccgcctcgcg cctccaggac ggagaatacg gccttggctg
     2041 cttggtcgga ggcgcctaca cttacggcgg cgggggctcc gcggccgcga cgggggctgg
     2101 aggagcgggg gcctatggta ctggaggtcc cggcggcccc ggaggcccgg caggcggcgg
     2161 cggcgcctgc agcatgggtc ctctgaccgg ctcctacaac gtgaacatgg ccttggcagg
     2221 cggccccggt cctggcggcg gcggcggcag cagcggcggt gccggggcac tcagcgctgc
     2281 gggggtaatc cgggtgccgg cacacaggcc gctcgccgga gccgtggccc acccccagcc
     2341 cctggccacc ggcttgccca ccgtgccctc tgtgcctgcc atgccgggcg tcaacaacct
     2401 cactggcctc accttcccct ggatggagag taaccgcaga tacacaaagg acaggttcac
     2461 aggtcacccc tatcagaacc ggacgccccc caagaagaag aagccgcgca cgtccttcac
     2521 acgcctgcag atctgcgagc tggagaagcg cttccaccgc cagaagtacc tggcctcggc
     2581 cgagcgcgcc gccctggcca aggcgctcaa aatgaccgat gcgcaggtca aaacctggtt
     2641 ccagaaccgg cggacaaagt ggagacggca gactgcggag gaacgggagg ccgagaggca
     2701 gcaagcgaac cgcatcctcc tgcagttgca gcaggaggcc ttccagaaga gcctggcaca
     2761 gccgctgccc gctgaccctc tgtgcgtgca caactcgtcg ctcttcgccc tgcagaatct
     2821 gcagccgtgg tctgatgact cgaccaaaat cactagcgtc acgtcggtgg cgtcggcctg
     2881 cgaggaccca gctttcttgt acaaagtggt gacgtaagct aggggcccgt ttaaacccgc
     2941 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3001 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3061 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3121 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3181 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3241 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3301 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3361 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3421 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     3481 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
     3541 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     3601 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     3661 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     3721 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     3781 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     3841 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     3901 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     3961 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     4021 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     4081 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     4141 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     4201 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     4261 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     4321 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     4381 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     4441 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     4501 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     4561 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4621 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4681 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4741 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4801 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4861 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4921 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4981 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5041 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5101 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5161 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5221 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5281 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5341 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5401 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5461 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5521 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5581 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5641 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5701 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5761 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5821 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5881 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5941 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6001 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6061 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6121 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6181 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6241 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6301 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6361 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6421 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6481 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6541 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6601 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6661 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6721 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6781 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6841 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6901 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6961 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     7021 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     7081 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     7141 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     7201 ttcgcgatgt acgggccaga tatacgcgtt
//
