LOCUS       sequence-476168-        6710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating Flp-In cell lines allowing dox-inducible 
            mammalian expression of 3xFlag-tagged full-length NRF1 isoform 
            mutated to be resistant to NRF1 siRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6710)
  TITLE     A Scalable Exon Deletion Platform Coupled To Single-Cell 
            Transcriptomics Reveals Exons Shaping Gene Expression Dynamics
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6710)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA5-3xFLAG-NRF1-FL_WobbleMut
FEATURES             Location/Qualifiers
     source          1..6710
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1034..1057
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1058..1082
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2596..2620)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2662..2679)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2668..2892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3025..3044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3176..3223
                     /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             3231..4253
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4383..4504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4420..4439)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4474..4493
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4553..4569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4553..4569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4566..4588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4577..4593
                     /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(4601..4607,4608..4625,4626..4631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4646..4667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4784..4801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4955..5543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5035..5054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5714..6505,6506..6574))
                     /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     6337..6356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6575..6679)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttccaccat ggactacaaa gaccatgacg gtgatacata
     1021 tgatcatgac atcgattaca aggatgacga tgacaagaca agtttgtaca aaaaagcagg
     1081 cttcgaggaa cacggagtga cccaaaccga acatatggct accatagaag cacatgcagt
     1141 ggcccagcaa gtgcagcagg tccatgtggc tacttacacc gagcatagta tgctgagtgc
     1201 tgatgaagac tcgccttctt ctcccgagga cacctcttac gatgactcag atatactcaa
     1261 ctccacagca gctgatgagg tgacagctca tctggcagct gcaggtcctg tgggaatggc
     1321 cgctgctgct gctgtggcaa caggaaagaa acggaaacgg cctcatgtat ttgagtctaa
     1381 tccatctatc cggaagaggc aacaaacacg tttgcttcgg aaacttcgag ccacgttaga
     1441 tgaatatact actcgtgtgg gacagcaagc catcgtgctg tgcattagcc cctccaaacc
     1501 taaccctgtc tttaaagtgt ttggtgcagc acctttggag aatgtggtgc gtaagtacaa
     1561 gagcatgatc ctggaagacc tggagtctgc tctggcagaa cacgcccctg cgccacagga
     1621 ggttaactca gaactgccgc ctctcaccat cgacggaatt ccagtctctg tggacaaaat
     1681 gacccaggcc cagcttcggg catttatccc agagatgctc aagtactcta caggtcgggg
     1741 aaaaccaggc tgggggaaag aaagctgcaa gcccatctgg tggcctgaag atatcccctg
     1801 ggcaaatgtc cggagtgatg tccgcacaga agagcaaaag cagagggttt catggaccca
     1861 ggcactacgg accatagtta aaaactgtta taaacagcat gggcgggaag accttttgta
     1921 tgcctttgaa gatcagcaaa cgcaaacaca ggccacagcc acacatagta tagctcatct
     1981 tgtaccatca cagactgtag tccagacttt tagtaaccct gatggcactg tctcacttat
     2041 ccaggttggt acgggggcaa cagtagccac attggctgat gcttcagaat tgccaaccac
     2101 ggtcaccgtt gcccaagtga attattctgc cgtggctgat ggagaggtgg aacaaaattg
     2161 ggccacgtta cagggaggtg agatgaccat ccagacgacg caagcatcag aggccaccca
     2221 ggcggtggca tcgttggcag aggccgcagt ggcagcttct caggagatgc agcagggagc
     2281 tacagtcact atggcgctta acagcgaagc tgccgcccat gctgtcgcca ccctggctga
     2341 ggccacctta caaggtgggg gacagatcgt cttgtctggg gaaaccgcag cagccgtcgg
     2401 agcacttact ggagtccaag atgctaatgg cctggtccag atccctgtga gcatgtacca
     2461 gactgtggtg accagcctcg cccagggcaa cggaccagtg caggtggcca tggcccctgt
     2521 gaccaccagg atatcagaca gcgcagtcac catggacggc caagctgtgg aggtggtgac
     2581 attggaacag tgataaccca gctttcttgt acaaagtggt ctgactcgag tctagagggc
     2641 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2701 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2761 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2821 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2881 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2941 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3001 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3061 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3121 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt acctagaagt
     3181 tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc aaaaagcctg
     3241 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     3301 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     3361 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     3421 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg
     3481 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     3541 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     3601 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     3661 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     3721 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     3781 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     3841 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     3901 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     3961 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     4021 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     4081 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     4141 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     4201 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac
     4261 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4321 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4381 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4441 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4501 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4741 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4801 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4861 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4921 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4981 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5041 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5101 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5161 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5221 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5281 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5341 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5401 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5461 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5521 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5581 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5641 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5701 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5761 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5821 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5881 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5941 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6001 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6061 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6121 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6181 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6241 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6301 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6361 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6421 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6481 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6541 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6601 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6661 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
