LOCUS       sequence-462188-        8066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG-tagged BACH2 (H. sapiens) in human cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8066)
  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 8066)
  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 FLAG-BACH2
FEATURES             Location/Qualifiers
     source          1..8066
                     /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             923..946
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             962..985
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1001..1024
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1061..1084
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3657..3674)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3663..3887
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3933..4361
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4020..4039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4230..4251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4370..4390)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4375..4704
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4555..4690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4617..4636
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4771..5565
                     /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(4825..4844)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5435..5454
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5739..5860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5776..5795)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5830..5849
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5909..5925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5909..5925)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5922..5944)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5933..5949
                     /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(5957..5963,5964..5981,5982..5987))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6002..6023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6140..6157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6311..6899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6391..6410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7070..7861,7862..7930))
                     /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     7693..7712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7931..8035)
                     /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 gctggctagc
      901 gtttaaactt aagcttacca tggactacaa ggacgacgat gacaaggtac caggatctat
      961 ggattataaa gatgatgatg ataaagggtc ggccgccagc tggagccacc ctcagttcga
     1021 gaagggagga ggaagcggcg gaggcagcgg aggaggaagc tggagccacc cgcagttcga
     1081 gaaaggagct ggatcctctg tggatgagaa gcctgactcc cccatgtatg tgtatgagtc
     1141 cacagtccac tgcaccaaca tcctcctggg cctcaatgac cagcggaaaa aggatattct
     1201 ctgtgacgtg actttgatcg tggagaggaa ggagttccgg gcccaccggg ctgtgctggc
     1261 cgcatgcagt gaatattttt ggcaggcgct ggttggacag acaaaaaatg atttggtggt
     1321 cagcttgcct gaggaggtca cagccagggg ctttgggccg ctgttacagt ttgcctacac
     1381 tgccaagctg ttactcagca gagaaaacat ccgcgaggtc atccgctgtg ctgagttcct
     1441 gcgcatgcac aacctggagg actcctgctt cagcttcctg cagacccagc tcctgaacag
     1501 tgaggatggc ctgtttgtgt gccggaagga tgctgcgtgc cagcgcccac acgaggactg
     1561 cgagaactct gcaggagagg aggaggatga agaggaggag acgatggatt cagagacggc
     1621 caagatggct tgccccaggg accagatgct tccagagccc atcagctttg aggccgccgc
     1681 catccccgta gcagagaagg aagaagccct gctgcccgag cctgacgtgc ccacagacac
     1741 caaggagagc tcagaaaagg acgcgttaac gcagtacccc agatacaaga aataccagct
     1801 tgcatgtacc aagaatgtct ataatgcatc atcacacagt acctcaggtt ttgcaagcac
     1861 attccgggaa gataactcta gcaacagcct caagccgggg cttgccaggg ggcagattaa
     1921 aagtgagccg cccagtgaag agaatgagga agagagcatc acgctctgcc tgtctggaga
     1981 tgagcctgac gccaaggaca gagcggggga tgtcgagatg gaccggaaac agcccagccc
     2041 tgcccctacc cccacggccc cagctggggc cgcctgcctg gagagatcca ggagcgtggc
     2101 ctcgccctcc tgcttaaggt ctctgttcag cataacgaaa agtgtggagc tgtctggcct
     2161 gcccagtaca tctcagcagc actttgccag gagtccagcc tgcccttttg acaaggggat
     2221 cactcagggt gaccttaaaa ctgactacac ccctttcaca gggaattatg gacagcccca
     2281 cgtgggccag aaggaggtgt ccaacttcac catggggtcg cccctcaggg ggcctgggtt
     2341 ggaggctctc tgtaaacagg agggagagct ggaccggagg agcgtgatct tctcctccag
     2401 cgcttgtgac caagtgagca cctcggtgca ttcttattct ggggtgagca gtttggacaa
     2461 agacctctct gagccggtgc caaagggtct gtgggtggga gccggccagt ccctccccag
     2521 ctcgcaggcc tactcccacg gtgggctgat ggccgaccac ttgccaggaa ggatgcggcc
     2581 caacaccagc tgcccggtgc caatcaaagt ctgccctcgc tcacccccct tggagaccag
     2641 gaccaggact tccagctcct gctcttccta ttcctacgcg gaggacggga gcgggggctc
     2701 accctgcagc ctccctctct gtgagttctc ctcctcgccc tgttcccagg gagccagatt
     2761 ccttgccaca gaacatcagg aaccaggcct gatgggagat ggaatgtaca accaagtgcg
     2821 gccccaaatt aaatgtgagc agtcttatgg aaccaactcc agtgacgaat ccggatcgtt
     2881 ctcggaagca gacagtgagt cgtgtcctgt gcaggacagg ggccaggagg taaaacttcc
     2941 ttttcctgta gatcaaatca cagatcttcc aaggaacgat ttccagatga tgattaaaat
     3001 gcacaagcta acctcagaac agttagagtt tattcatgat gtccgacggc gcagcaagaa
     3061 ccgcatcgcg gcccagcgct gccgcaaaag gaaactggac tgtattcaga atttagaatg
     3121 tgaaatccgc aaattggtgt gtgagaaaga gaaactgttg tcagagagga atcaactgaa
     3181 agcatgcatg ggggaactgt tggacaactt ctcctgcctt tcccaggaag tttgccgaga
     3241 catccagagc cccgagcaga tccaggccct gcatcggtat tgccctgtcc tcagacccat
     3301 ggacttgccc acggcctcca gtattaaccc tgcgcccttg ggtgctgagc agaacattgc
     3361 ggcctcccaa tgcgcagtgg gggaaaacgt gccctgctgc ttggagccag gcgcggctcc
     3421 ccccggaccc ccctgggcac ccagcaacac ctccgagaat tgtacctctg ggaggagact
     3481 agaaggcact gacccgggaa ccttctcaga gagaggacct cctcttgaac ccaggagcca
     3541 aacagtgacc gtggacttct gccaggaaat gactgataag tgtacaactg acgaacagcc
     3601 caggaaagat tatacctagc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3661 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3721 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3781 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3841 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3901 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3961 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4021 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4081 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4141 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4201 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4261 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4321 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4381 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4441 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4501 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4561 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4621 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4681 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4741 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4801 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4861 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4921 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4981 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5041 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5101 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5161 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5221 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5281 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5341 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5401 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5461 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5521 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5581 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5641 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5701 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5761 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5821 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5881 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5941 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6001 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6061 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6121 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6181 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6241 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6721 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8041 ttccccgaaa agtgccacct gacgtc
//
