LOCUS       sequence-333511-        7506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PAX6 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7506)
  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 7506)
  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_PAX6
FEATURES             Location/Qualifiers
     source          1..7506
                     /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(3162..3186)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3224..3241)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3230..3454
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3738..3785
                     /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             3793..4815
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3793..4815
                     /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    4945..5066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4982..5001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5036..5055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5115..5131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5115..5131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5128..5150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5139..5155
                     /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(5163..5169,5170..5187,5188..5193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5208..5229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5346..5363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5517..6105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5597..5616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6276..7067,7068..7136))
                     /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     6899..6918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7137..7241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7316..7335)
                     /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 caccatgcag aacagtcaca gcggagtgaa
     1921 tcagctcggt ggtgtctttg tcaacgggcg gccactgccg gactccaccc ggcagaagat
     1981 tgtagagcta gctcacagcg gggcccggcc gtgcgacatt tcccgaattc tgcaggtgtc
     2041 caacggatgt gtgagtaaaa ttctgggcag gtattacgag actggctcca tcagacccag
     2101 ggcaatcggt ggtagtaaac cgagagtagc gactccagaa gttgtaagca aaatagccca
     2161 gtataagcgg gagtgcccgt ccatctttgc ttgggaaatc cgagacagat tactgtccga
     2221 gggggtctgt accaacgata acataccaag cgtgtcatca ataaacagag ttcttcgcaa
     2281 cctggctagc gaaaagcaac agatgggcgc agacggcatg tatgataaac taaggatgtt
     2341 gaacgggcag accggaagct ggggcacccg ccctggttgg tatccgggga cttcggtgcc
     2401 agggcaacct acgcaagatg gctgccagca acaggaagga gggggagaga ataccaactc
     2461 catcagttcc aacggagaag attcagatga ggctcaaatg cgacttcagc tgaagcggaa
     2521 gctgcaaaga aatagaacat cctttaccca agagcaaatt gaggccctgg agaaagagtt
     2581 tgagagaacc cattatccag atgtgtttgc ccgagaaaga ctagcagcca aaatagatct
     2641 acctgaagca agaatacagg tatggttttc taatcgaagg gccaaatgga gaagagaaga
     2701 aaaactgagg aatcagagaa gacaggccag caacacacct agtcatattc ctatcagcag
     2761 tagtttcagc accagtgtct accaaccaat tccacaaccc accacaccgg tttcctcctt
     2821 cacatctggc tccatgttgg gccgaacaga cacagccctc acaaacacct acagcgctct
     2881 gccgcctatg cccagcttca ccatggcaaa taacctgcct atgcaacccc cagtccccag
     2941 ccagacctcc tcatactcct gcatgctgcc caccagccct tcggtgaatg ggcggagtta
     3001 tgatacctac acccccccac atatgcagac acacatgaac agtcagccaa tgggcacctc
     3061 gggcaccact tcaacaggac tcatttcccc tggtgtgtca gttccagttc aagttcccgg
     3121 aagtgaacct gatatgtctc aatactggcc aagattacag gacccagctt tcttgtacaa
     3181 agtggtgacg taagctaggg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3241 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3301 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3361 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3421 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3481 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3721 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     3781 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     3841 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     3901 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     3961 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     4021 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     4081 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     4141 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     4201 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     4261 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     4321 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     4381 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     4441 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     4501 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     4561 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     4621 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     4681 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     4741 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     4801 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4861 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4921 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4981 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5041 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5101 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7261 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     7321 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7381 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     7441 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     7501 cgcgtt
//
