LOCUS       sequence-462193-        7751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N-terminal tagged 2xFLAG-2xSTREP-BACH1 (H. sapiens) 
            with Cys122 to Glu point mutation in human cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7751)
  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 7751)
  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 FFSS-BACH1 C122E
FEATURES             Location/Qualifiers
     source          1..7751
                     /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(3342..3359)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3348..3572
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3618..4046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3705..3724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3915..3936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4055..4075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4060..4389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4240..4375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4302..4321
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4456..5250
                     /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(4510..4529)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5120..5139
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5424..5545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5461..5480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5515..5534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5594..5610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5594..5610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5607..5629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5618..5634
                     /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(5642..5648,5649..5666,5667..5672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5687..5708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5825..5842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5996..6584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6076..6095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6755..7546,7547..7615))
                     /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     7378..7397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7616..7720)
                     /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 ggatcctctc tgagtgagaa ctcggttttt gcctatgaat cttctgtgca
     1141 tagcaccaat gttttactca gccttaatga ccagcggaag aaagatgtgc tgtgcgatgt
     1201 caccatcttt gtggagggac agcggttccg cgctcaccgg tccgtgctgg cggcatgcag
     1261 cagttacttc cactcaagaa tcgtaggcca ggctgatgga gagctgaaca ttactcttcc
     1321 agaagaggtg acagttaaag gatttgaacc tttaattcag tttgcctaca ctgctaaact
     1381 gattttaagt aaagagaatg tggatgaagt gtgcaaatgt gtggagtttt taagtgtaca
     1441 taatattgag gaatccgaat ttcagtttct gaaatttaag tttttggact ccactgcaga
     1501 ccagcaagaa tgcccaagaa aaaaatgctt ttcatcacac tgtcagaaaa cagaccttaa
     1561 actttcactt ttggaccaga gggatctaga aactgatgaa gtggaggaat ttctggaaaa
     1621 taaaaatgtt cagactcctc agtgtaaact ccgcaggtat caaggaaatg caaaagcctc
     1681 acctcctcta caagacagtg ccagtcagac atatgagtcc atgtgcttag agaaggatgc
     1741 tgctctggcc ttgccttctt tatgccccaa atacagaaaa ttccaaaaag catttggaac
     1801 tgacagagtc cgtactgggg aatctagtgt caaagacatt catgcttctg ttcagccaaa
     1861 tgaaaggtct gaaaatgaat gcctgggagg agtcccggag tgtagagatt tgcaggtgat
     1921 gttaaaatgt gacgaaagta aattagcaat ggaacctgaa gaaacgaaga aagatcctgc
     1981 ttctcagtgc ccaactgaaa aatcagaagt gactcctttc ccccacaatt cttccataga
     2041 ccctcatgga ctttattctt tgtctctttt acacacatat gaccaatatg gtgacttgaa
     2101 ttttgctggt atgcaaaaca caacagtgtt aacagaaaag cctttgtcag gtacagacgt
     2161 ccaagaaaaa acatttggtg aaagtcagga tttacctttg aaatccgact tgggcaccag
     2221 ggaagatagt agtgttgcat ctagtgatag gagtagtgtg gagcgagaag tggcagaaca
     2281 cctagcaaaa ggcttctgga gtgacatttg cagcacggac actccttgcc aaatgcagtt
     2341 atcacctgct gtggccaaag atggctcaga acagatctca cagaaacggt ctgagtgtcc
     2401 gtggttaggt atcaggatta gtgagagccc agaaccaggt caaaggactt tcacaacatt
     2461 aagttctgtc aactgccctt ttataagtac tctgagtact gaaggctgtt caagcaattt
     2521 ggaaattgga aacgatgatt atgtttcaga accccagcaa gaaccttgcc catatgcttg
     2581 tgtcattagc ttgggagacg actctgagac ggacaccgaa ggagacagtg aatcctgttc
     2641 agccagagaa caagaatgtg aggtaaaact gccattcaat gcacaacgga taatttcact
     2701 gtctcgaaat gattttcagt ccttgttgaa aatgcacaag cttactccag aacagctgga
     2761 ttgtatccat gatattcgaa gaagaagtaa aaacagaatt gctgcacagc gctgtcgcaa
     2821 gagaaaactt gactgtatac agaatcttga atcagaaatt gagaagctgc aaagtgaaaa
     2881 ggagagcttg ttgaaggaaa gagatcacat tttgtcaact ctgggtgaga caaagcagaa
     2941 cctaactgga ctttgccaga aagtttgtaa agaagcagct ctgagtcaag aacaaataca
     3001 gatactcgcc aagtactcag ctgcagattg cccactttca tttttaattt ctgaaaaaga
     3061 taaaagtact cctgatggtg aactggcgtt accatcaatt ttcagtttat ctgaccggcc
     3121 tccagcagtg ctgcctccct gtgccagagg aaacagtgag cctggctacg cgcgagggca
     3181 ggagtcccag cagatgtcca cagccacctc tgagcaagct gggcctgcgg aacagtgtcg
     3241 tcagagtggt gggatctcag atttctgtca gcagatgact gataaatgta ctactgatga
     3301 gtaactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3361 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgcaac
     3421 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3481 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3541 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3601 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3661 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3721 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3781 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3841 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3901 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3961 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4021 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4081 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4141 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4201 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4261 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4321 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4381 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4441 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4501 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4561 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4621 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4681 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4741 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4801 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4861 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4921 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4981 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5041 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5101 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5161 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5221 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5281 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5341 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5401 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5461 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5521 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5581 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5641 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5701 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5761 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5821 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5881 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5941 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6001 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6061 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6121 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6181 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6241 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6301 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6361 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6421 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6481 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6541 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6601 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6661 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6721 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6781 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6841 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6901 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6961 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7021 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7081 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7141 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7201 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7261 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7321 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7381 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7441 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7501 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7561 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7621 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7681 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7741 cacctgacgt c
//
