LOCUS       sequence-338259-        7766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG tagged human RBPJ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7766)
  AUTHORS   Zhang T, Guo L, Creighton CJ, Lu Q, Gibbons DL, Yi ES, Deng B, 
            Molina JR, Sun Z, Yang P, Yang Y
  TITLE     A genetic cell context-dependent role for ZEB1 in lung cancer.
  JOURNAL   Nat Commun. 2016 Jul 26;7:12231. doi: 10.1038/ncomms12231.
  PUBMED    27456471
REFERENCE   2  (bases 1 to 7766)
  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: "Nat 
            Commun."; date: "2016-07-26"; volume: "7:12231. doi"; pages: " 
            10.1038/ncomms12231"
COMMENT     Sequence Label: PCMV10-RBPJ
FEATURES             Location/Qualifiers
     source          1..7766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2208..2830
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2407..2426)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2854..2874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2872..3169
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3223..4017
                     /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(3277..3296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3887..3906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4669..4803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4706..4725)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4760..4779
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4808..4827)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4809..4827)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4841..4857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4841..4857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4854..4876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4865..4881
                     /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(4889..4895,4896..4913,4914..4919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4934..4955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5072..5089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5243..5831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5323..5342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6002..6793,6794..6862))
                     /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     6625..6644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6863..6967)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6994..7449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7081..7100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7291..7312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7575..7597
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7589..7606
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7590..7606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccgc gatggaccac acggagggct cgcccgcgga
      721 ggagccgcct gcgcatgttc catcgcctgg gaaatttggt gagcggcctc cacctaaacg
      781 acttactagg gaagctatgc gaaattattt aaaagagcga ggggatcaaa cagtacttat
      841 tcttcatgca aaagttgcac agaagtcata tggaaatgaa aaaaggtttt tttgcccacc
      901 tccttgtgta tatcttatgg gcagtggatg gaagaaaaaa aaagaacaaa tggaacgcga
      961 tggttgttct gaacaagagt ctcaaccgtg tgcatttatt gggataggaa atagtgacca
     1021 agaaatgcag cagctaaact tggaaggaaa gaactattgc acagccaaaa cattgtatat
     1081 atctgactca gacaagcgaa agcacttcat gttgtctgta aagatgttct atggcaacag
     1141 tgatgacatt ggtgtgttcc tcagcaagcg gataaaagtc atctccaaac cttccaaaaa
     1201 gaagcagtca ttgaaaaatg ctgacttatg cattgcctca ggaacaaagg tggctctgtt
     1261 taatcgacta cgatcccaga cagttagtac cagatacttg catgtagaag gaggtaattt
     1321 tcatgccagt tcacagcagt ggggagcctt ttttattcat ctcttggatg atgatgaatc
     1381 agaaggagaa gaattcacag tccgagatgg ctacatccat tatggacaaa cagtcaaact
     1441 tgtgtgctca gttactggca tggcactccc aagattgata attaggaaag ttgataagca
     1501 gaccgcatta ttggatgcag atgatcctgt gtcacaactc cataaatgtg cattttacct
     1561 taaggataca gaaagaatgt atttgtgcct ttctcaagaa agaataattc aatttcaggc
     1621 cactccatgt ccaaaagaac caaataaaga gatgataaat gatggcgctt cctggacaat
     1681 cattagcaca gataaggcag agtatacatt ttatgaggga atgggccctg tccttgcccc
     1741 agtcactcct gtgcctgtgg tagagagcct tcagttgaat ggcggtgggg acgtagcaat
     1801 gcttgaactt acaggacaga atttcactcc aaatttacga gtgtggtttg gggatgtaga
     1861 agctgaaact atgtacaggt gtggagagag tatgctctgt gtcgtcccag acatttctgc
     1921 attccgagaa ggttggagat gggtccggca accagtccag gttccagtaa ctttggtccg
     1981 aaatgatggg atcatttatt ccaccagcct tacctttacc tacacaccag aaccagggcc
     2041 gcggccacat tgcagtgcag caggagcaat ccttcgagcc aattcaagcc aggtgccccc
     2101 taacgaatca aacacaaaca gcgagggaag ttacacaaac gccagcacaa attcaaccag
     2161 tgtcacatca tctacagcca cagtggtatc ctaatctaga ggatcccggg tggcatccct
     2221 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     2281 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     2341 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     2401 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     2461 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     2521 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     2581 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     2641 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac
     2701 tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg
     2761 gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc
     2821 ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa
     2881 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2941 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3001 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3061 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3121 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggaattgatc
     3181 agcttgggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3241 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3301 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3361 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3421 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3481 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3541 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3601 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3661 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3721 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3781 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3841 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3901 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3961 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4021 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4081 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4141 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc gggctcgatc
     4201 ccctcgcgag ttggttcagc tgctgcctga ggctggacga cctcgcggag ttctaccggc
     4261 agtgcaaatc cgtcggcatc caggaaacca gcagcggcta tccgcgcatc catgcccccg
     4321 aactgcagga gtggggaggc acgatggccg ctttggtcga cccggacggg acgctcctgc
     4381 gcctgataca gaacgaattg cttgcaggca tctcatgagt gtgtcttccc gttttccgcc
     4441 tgaggtcact gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc tgctcaccac
     4501 ccactcgcca agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4561 ccgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4621 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4681 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4741 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4801 atcaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt
     4861 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4921 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4981 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5641 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6961 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     7021 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     7081 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     7141 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7201 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7261 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7321 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     7381 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     7441 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7501 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7561 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctgatcta
     7621 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
     7681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     7741 ctcatgtcca atatgaccgc catgtt
//
