LOCUS       sequence-372405-        7635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human PAX2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7635)
  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 7635)
  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_PAX2
FEATURES             Location/Qualifiers
     source          1..7635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        123..502
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        503..706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     657..677
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    708..726
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    729..747
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     757..781
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(949..1299,1300..1302,1303..1911)
                     /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             1912..1938
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1969..1992
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2029..2052
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    2065..2089
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3413..3437)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3475..3492)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3481..3705
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3838..3857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3989..4036
                     /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             4044..5066
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4044..5066
                     /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    5196..5317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5233..5252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5287..5306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5366..5382)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5366..5382)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5379..5401)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5390..5406
                     /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(5414..5420,5421..5438,5439..5444))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5459..5480
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5597..5614)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5768..6356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5848..5867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6527..7318,7319..7387))
                     /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     7150..7169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7388..7492)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7567..7586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
       61 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      121 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      181 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      241 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      301 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      361 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      421 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      481 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      541 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      601 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      661 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
      721 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
      781 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
      841 cctccggact ctagcgttta aacttaagct tggtaccgag ctcggatcca ctagtccagt
      901 gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagaccat gaaggacaac
      961 accgtgcccc tgaagctgat cgccctgctg gccaacggcg agttccactc tggcgagcag
     1021 ctgggcgaga cactgggcat gagcagagcc gccatcaaca agcacatcca gaccctgcgg
     1081 gactggggcg tggacgtgtt cacagtgccc ggcaagggct acagcctgcc cgagcccatc
     1141 cagctgctga acgccaagca gatcctgggc cagctggatg gcggatctgt ggccgtgctg
     1201 cccgtgatcg acagcaccaa ccagtatctg ctcgaccgga tcggcgagct gaagtccggc
     1261 gacgcctgta tcgccgagta ccagcaggcc ggcagaggcg gcaggggcag aaagtggttc
     1321 agccccttcg gcgccaacct gtacctgagc atgttctgga gactggaaca gggcccagcc
     1381 gccgctattg gcctgagcct ggtcatcggc atcgtgatgg ccgaggtgct gcggaagctg
     1441 ggcgccgaca aagtgcgcgt gaagtggccc aacgacctgt atctgcagga ccggaagctg
     1501 gccggcatcc tggtggaact gaccggcaaa acaggcgacg ccgcccagat cgtgatcggc
     1561 gctggcatca acatggccat gcggcgggtg gaagagagcg tggtcaacca gggctggatc
     1621 accctgcagg aagccggcat caacctggac cggaacaccc tggccgccat gctgatcaga
     1681 gagctgagag ccgccctgga actgttcgag caggaaggcc tggcccccta cctgagcaga
     1741 tgggagaagc tggacaactt catcaaccgg cccgtgaaac tgatcatcgg cgacaaagag
     1801 atcttcggca tcagccgggg catcgacaag cagggggccc tgctcctgga acaggacggc
     1861 atcatcaagc cctggatggg cggcgagatc agcctgcgga gcgccgagaa gtacccatac
     1921 gatgttcctg actatgccgg taccgagctc ggatccacca tggctagctg gagccacccg
     1981 cagttcgaga aaggtggagg ttccggaggt ggatcgggag gtggatcgtg gagccacccg
     2041 cagttcgaaa aagcggccga tatcacaagt ttgtacaaaa aagcagaagg gccgtcaagg
     2101 cccaccatgg atatgcactg caaagcagac cccttctccg cgatgcaccc agggcacggg
     2161 ggtgtgaacc agctcggggg ggtgtttgtg aacggccggc ccctacccga cgtggtgagg
     2221 cagcgcatcg tggagctggc ccaccagggt gtgcggccct gtgacatctc ccggcagctg
     2281 cgggtcagcc acggctgtgt cagcaaaatc ctgggcaggt actacgagac cggcagcatc
     2341 aagccgggtg tgatcggtgg ctccaagccc aaagtggcga cgcccaaagt ggtggacaag
     2401 attgctgaat acaaacgaca gaacccgact atgttcgcct gggagattcg agaccggctc
     2461 ctggccgagg gcatctgtga caatgacaca gtgcccagcg tctcttccat caacagaatc
     2521 atccggacca aagttcagca gcctttccac ccaacgccgg atggggctgg gacaggagtg
     2581 accgcccctg gccacaccat tgttcccagc acggcctccc ctcctgtttc cagcgcctcc
     2641 aatgacccag tgggatccta ctccatcaat gggatcctgg ggattcctcg ctccaatggt
     2701 gagaagagga aacgtgatga agttgaggta tacactgatc ctgcccacat tagaggaggt
     2761 ggaggtttgc atctggtctg gactttaaga gatgtgtctg agggctcagt ccccaatgga
     2821 gattcccaga gtggtgtgga cagtttgcgg aagcacttgc gagctgacac cttcacccag
     2881 cagcagctgg aagctttgga tcgggtcttt gagcgtcctt cctaccctga cgtcttccag
     2941 gcatcagagc acatcaaatc agaacagggg aacgagtact ccctcccagc cctgacccct
     3001 gggcttgatg aagtcaagtc gagtctatct gcatccacca accctgagct gggcagcaac
     3061 gtgtcaggca cacagacata cccagttgtg actggtcgtg acatggcgag caccactctg
     3121 cctggttacc cccctcacgt gccccccact ggccagggaa gctaccccac ctccaccctg
     3181 gcaggaatgg tgcctgggag cgagttctcc ggcaacccgt acagccaccc ccagtacacg
     3241 gcctacaacg aggcttggag attcagcaac cccgccttac taatgccgcc ccccggtccg
     3301 cccctgccgc tgctgccgct gcctatgacc gccactagtt accgcgggga ccacatcaag
     3361 cttcaggccg acagcttcgg cctccacatc gtccccgtct caggcctcat gggcccagct
     3421 ttcttgtaca aagtggtgac gtaagctagg ggcccgttta aacccgctga tcagcctcga
     3481 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3541 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3601 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3661 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3721 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3781 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3841 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3901 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3961 ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag
     4021 aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     4081 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     4141 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     4201 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     4261 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     4321 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     4381 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     4441 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     4501 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     4561 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     4621 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     4681 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     4741 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     4801 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     4861 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     4921 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     4981 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     5041 gcactcgtcc gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct
     5101 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5161 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5221 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5281 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5341 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5401 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5461 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5521 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5581 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5641 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5701 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5761 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5821 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5881 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5941 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6001 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6061 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6121 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6181 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6241 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6301 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6361 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6421 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6481 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6541 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6601 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6661 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6721 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6781 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6841 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6901 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6961 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7021 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7081 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7141 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7201 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7261 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7321 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7381 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7441 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7501 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     7561 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     7621 gttggaggtc gctga
//
