LOCUS       sequence-471110-        6139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express BirA and HA tagged TULP3 transiently via transfection in 
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6139)
  AUTHORS   Jiang Y, Xun Y, Zhang Z
  TITLE     Central regulation of feeding and body weight by ciliary GPR75.
  JOURNAL   J Clin Invest. 2024 Aug 13;134(19):e182121. doi: 10.1172/JCI182121.
  PUBMED    39137039
REFERENCE   2  (bases 1 to 6139)
  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: "J Clin 
            Invest."; date: "2024-08-13"; volume: "134(19):e182121. doi"; pages:
            " 10.1172/JCI182121"
COMMENT     Sequence Label: pCMV-BirA-HA-Tulp3
FEATURES             Location/Qualifiers
     source          1..6139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1204..1222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1260..1282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1382..1401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1595..1617
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1609..1626
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1610..1626
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        1653..1956
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1957..2160
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2111..2131
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2157..2181
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          2298..2394
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(2452..2802,2803..2805,2806..3414)
                     /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             3445..3471
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4904..5038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4928..4947)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4982..5001
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5149..5165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5149..5165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5162..5184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5173..5189
                     /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(5197..5203,5204..5221,5222..5227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5242..5263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5380..5397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5551..6139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5631..5650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1201 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     1261 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1321 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     1381 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1441 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     1501 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1561 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1621 gccagtgagt tcgagcttgc atgcctgcag gtcgttacat aacttacggt aaatggcccg
     1681 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     1741 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     1801 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1861 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1921 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     1981 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2041 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     2101 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     2161 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     2221 agacaccggg accgatccag cctccggact ctagaggatc cggtactaga ggaactgaaa
     2281 aaccagaaag ttaactggta agtttagtct ttttgtcttt tatttcaggt cccggatccg
     2341 gtggtggtgc aaatcaaaga actgctcctc agtggatgtt gcctttactt ctaggcctgt
     2401 acggaagtgt tacttctgct ctaaaagctg cggaattgta cccgcgccac catgaaggac
     2461 aacaccgtgc ccctgaagct gatcgccctg ctggccaacg gcgagttcca cagcggcgag
     2521 cagctgggcg agaccctggg catgagccgg gccgccatca acaagcacat ccagaccctg
     2581 cgggactggg gcgtggacgt gttcaccgtg cccggcaagg gctacagcct gcccgagccc
     2641 atccagctgc tgaacgccaa gcagatcctg ggccagctgg acggcggcag cgtggccgtg
     2701 ctgcccgtga tcgacagcac caaccagtac ctgctggacc ggatcggcga gctgaagagc
     2761 ggcgacgcct gcatcgccga gtaccagcag gccggccggg gcggccgggg ccggaagtgg
     2821 ttcagcccct tcggcgccaa cctgtacctg agcatgttct ggcggctgga gcagggcccc
     2881 gccgccgcca tcggcctgag cctggtgatc ggcatcgtga tggccgaggt gctgcggaag
     2941 ctgggcgccg acaaggtgcg ggtgaagtgg cccaacgacc tgtacctgca ggaccggaag
     3001 ctggccggca tcctggtgga gctgaccggc aagaccggcg acgccgccca gatcgtgatc
     3061 ggcgccggca tcaacatggc catgcggcgg gtggaggaga gcgtggtgaa ccagggctgg
     3121 atcaccctgc aggaggccgg catcaacctg gaccggaaca ccctggccgc catgctgatc
     3181 cgggagctgc gggccgccct ggagctgttc gagcaggagg gcctggcccc ctacctgagc
     3241 cggtgggaga agctggacaa cttcatcaac cggcccgtga agctgatcat cggcgacaag
     3301 gagatcttcg gcatcagccg gggcatcgac aagcagggcg ccctgctgct ggagcaggac
     3361 ggcatcatca agccctggat gggcggcgag atcagcctgc ggagcgccga gaagggggga
     3421 tctggccccg gcggcggcgc gccataccca tacgatgttc cagattacgc tcttatggcc
     3481 atggaggccc gaattcggat ggaggcggcg cgctgcgcac ccggaccccg cggggacagt
     3541 gcctttgacg atgagaccct gaggcttcgg cagctgaagc tggacaatca gagggcactg
     3601 ctggagaaga agcagagaaa gaagcgcctt gagccactca tggtacagcc aaacccagaa
     3661 gccaggctgc gaaggttaaa gccacgaggc agtgaggagc acacgcctct ggtggaccct
     3721 cagatgcctc gcagcgacgt catcctacac ggcatcgacg gtccagctgc tttccttaag
     3781 ccagaggctc aggatttgga aagcaagcct caggttctct ctgtgggctc ccctgcccca
     3841 gaagaaggca cggaagggag tgcagatggg gaaagccccg aggagacagc ccccaagcca
     3901 gaccttcagg agattctcca aaaacacggc atcttgagta gtgtgaacta tgacgaggag
     3961 cctgacaagg aggaagatga ggggggaaac ctcagctccc cgtcggctcg ctcagaggag
     4021 agtgctgcag ccagccagaa agcagcctcg gagacaggag cttccggtgt gacagcccag
     4081 cagggtgacg cccagcttgg agaagtggag aatttagagg actttgcgta tagccctgcc
     4141 cctcggggtg tcacggtgaa gtgtaaagta accagggata agaaaggcat ggaccgtggc
     4201 ctcttcccca cctactatat gcacctggaa cgggaggaga accgcaagat atttcttcta
     4261 gccggtagaa agcggaaaaa gagcaaaacg tccaactacc tggtctccac agacccgaca
     4321 gatttgtctc gtgaagggga aagttacatt ggcaaactca gatccaatct catggggacc
     4381 aagttcacag tgtatgacca tggtgtcaac ccagtcaagg cccagggtct ggtggagaag
     4441 gcccacaccc gccaggagct ggctgctatc tgttatgaaa caaatgtcct cggtttcaaa
     4501 ggtcccagga aaatgtctgt gatcatcccg gggatgaaca tgaaccatga gcgcatccct
     4561 ttcaggccac gaaatgaaca cgagagcttg ctttcaaaat ggcaaaacaa atccatggag
     4621 aacctgatcg agctgcacaa caaggccccg gtgtggaatg acgacaccca gtcctacgtg
     4681 ctgaactttc acggcagagt cacccaggca tccgtgaaaa acttccagat cgtccacggg
     4741 aatgaccctg actacatcgt catgcagttt ggacgcgtgg ctgatgacgt gttcacactg
     4801 gactataact acccactgtg cgcacttcaa gccttcgcca tcgggctgtc cagctttgac
     4861 agcaagctgg cgtgtgaatg actcgaggta ccgcggccgc ggggatccag acatgataag
     4921 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4981 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     5041 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc
     5101 ggatcctcta gagtcgatct gcaggcatgc tagcttggcg taatcatggt catagctgtt
     5161 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5221 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5281 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6121 cagaaaaaaa ggatctcaa
//
