LOCUS       sequence-462190-        6626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N-terminal tagged HA-FBXO22 (H. sapiens) in human cell
            lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6626)
  AUTHORS   Cao S, Garcia SF, Shi H, James EI, Kito Y, Shi H, Mao H, Kaisari S, 
            Rona G, Deng S, Goldberg HV, Ponce J, Ueberheide B, Lignitto L, 
            Guttman M, Pagano M, Zheng N
  TITLE     Recognition of BACH1 quaternary structure degrons by two F-box 
            proteins under oxidative stress.
  JOURNAL   Cell. 2024 Oct 30:S0092-8674(24)01188-7. doi: 
            10.1016/j.cell.2024.10.012.
  PUBMED    39504958
REFERENCE   2  (bases 1 to 6626)
  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: "Cell. 2024 
            Oct 30:S0092-8674(24)01188-7. doi: 10.1016/j.cell.2024.10.012."
COMMENT     Sequence Label: pcDNA3.1 HA-FBXO22
FEATURES             Location/Qualifiers
     source          1..6626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             910..936
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(2179..2197)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2180..2197)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2217..2234)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2223..2447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2493..2921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2580..2599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2790..2811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2930..2950)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2935..3264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3115..3250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3177..3196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3331..4125
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3385..3404)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3995..4014
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4299..4420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4336..4355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4390..4409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4469..4485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4469..4485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4482..4504)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4493..4509
                     /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(4517..4523,4524..4541,4542..4547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4562..4583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4700..4717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4871..5459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4951..4970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5630..6421,6422..6490))
                     /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     6253..6272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6491..6595)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
      901 gcctacccct acgacgtgcc cgactacgcc tccctcggat ctgagccggt aggctgctgc
      961 ggcgagtgcc gcggctcctc cgtagacccg cggagcacct tcgtgttgag taacctggcg
     1021 gaggtggtgg agcgtgtgct caccttcctg cccgccaagg cgttgctgcg ggtggcctgc
     1081 gtgtgccgct tatggaggga gtgtgtgcgc agagtattgc ggacccatcg gagcgtaacc
     1141 tggatctccg caggcctggc ggaggccggc cacctggagg ggcattgctt ggttcgcgtg
     1201 gtagcagagg agcttgagaa tgttcgcatc ttaccacata cagttcttta catggctgat
     1261 tcagaaactt tcattagtct ggaagagtgt cgtggccata agagagcaag gaaaagaact
     1321 agtatggaaa cagcacttgc ccttgagaag ctattcccca aacaatgcca agtccttggg
     1381 attgtgaccc caggaattgt agtgactcca atgggatcag gtagcaatcg acctcaggaa
     1441 atagaaattg gagaatctgg ttttgcttta ttattccctc aaattgaagg aataaaaata
     1501 caaccctttc attttattaa ggatccaaag aatttaacat tagaaagaca tcaactcact
     1561 gaagtaggtc ttttagataa ccctgaactt cgtgtggtcc ttgtctttgg ttataattgc
     1621 tgtaaggtgg gagccagtaa ttatctgcag caagtagtca gcactttcag tgatatgaat
     1681 atcatcttgg ctggaggcca ggtggacaac ctgtcatcac tgacttctga aaagaaccct
     1741 ctggatattg atgcctcggg tgtggttgga ctgtcattta gtggacaccg aatccagagt
     1801 gccactgtgc tcctcaacga ggacgtcagt gatgagaaga ctgctgaggc tgcgatgcag
     1861 cgcctcaaag cggccaacat tccagagcat aacaccattg gcttcatgtt tgcatgcgtt
     1921 ggcaggggct ttcagtatta cagagccaag gggaatgttg aggctgatgc atttagaaag
     1981 ttttttccta gtgttccctt attcggcttc tttggaaatg gagaaattgg atgtgatcgg
     2041 atagtcactg ggaactttat attgaggaaa tgtaatgagg taaaagatga tgatctgttt
     2101 catagctata caacaataat ggcactcata catctggggt catctaaata actcgagcat
     2161 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     2221 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2281 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2341 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2401 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2461 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2521 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2581 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2641 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2701 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2761 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2821 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2881 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2941 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3001 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3061 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3121 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3181 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3241 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3301 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3361 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3421 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3481 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3541 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3601 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3661 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3721 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3781 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3841 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3901 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3961 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4021 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4081 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4141 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4201 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4261 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4321 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4381 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4441 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4501 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4561 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4621 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4681 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4741 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4801 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5041 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5281 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6601 ttccccgaaa agtgccacct gacgtc
//
