LOCUS       sequence-424529-        8052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8052)
  AUTHORS   Lei K, Nimnual A, Zong WX, Kennedy NJ, Flavell RA, Thompson CB, 
            Bar-Sagi D, Davis RJ
  TITLE     The Bax subfamily of Bcl2-related proteins is essential for 
            apoptotic signal transduction by c-Jun NH(2)-terminal kinase.
  JOURNAL   Mol Cell Biol. 2002 Jul . 22(13):4929-42.
  PUBMED    12052897
REFERENCE   2  (bases 1 to 8052)
  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: "Mol Cell 
            Biol. 2002 Jul . 22(13):4929-42."
COMMENT     Sequence Label: pCDNA3 Flag MKK7B2Jnk1a1(APF)
FEATURES             Location/Qualifiers
     source          1..8052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        109..488
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        489..692
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     643..663
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     737..756
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        737..755
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             783..806
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(933..944)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(3479..3497)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3480..3497)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3517..3534)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3523..3747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3793..4221
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3880..3899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4090..4111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4230..4250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4235..4564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4415..4550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4477..4496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4631..5425
                     /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(4685..4704)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5295..5314
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5599..5720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5636..5655)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5690..5709
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5769..5785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5769..5785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5782..5804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5793..5809
                     /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(5817..5823,5824..5841,5842..5847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5862..5883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6000..6017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6171..6759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6251..6270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6930..7721,7722..7790))
                     /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     7553..7572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7791..7895)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7970..7989)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
       61 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      121 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      181 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      241 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      301 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      361 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      421 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      481 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      541 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      601 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      661 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      721 actggcttat cgaaattaat acgactcact atagggagac ccaagcttcg gcagccaaca
      781 tggactataa ggacgatgat gacaaaatgg cggcgtcctc cctggagcag aagctgtccc
      841 gcctggaagc caagctgaag caggagaacc gtgaggcccg caggaggatc gacctcaact
      901 tggatatcag cccacagcgg cccaggccca ccctgcaact cccactggcc aacgatgggg
      961 gcagccgctc accatcctca gagagctccc cacagcaccc tacacccccc acccggcccc
     1021 gccacatgct ggggctccca tcaaccttgt tcacaccgcg cagtatggag agcatcgaga
     1081 ttgaccagaa gctgcaggag atcatgaagc agacagggta cctgactatc gggggccagc
     1141 gttatcaggc agaaatcaat gacttggaga acttgggtga gatgggcagt ggtacctgtg
     1201 gtcaggtgtg gaagatgcgg ttccggaaga caggccacat cattgctgtt aagcaaatgc
     1261 ggcgctctgg gaacaaggaa gagaataagc gcattttgat ggacctggat gtagtactca
     1321 agagccatga ctgcccttac atcgttcagt gctttggcac cttcatcacc aacacagacg
     1381 tctttattgc catggagctc atgggcacat gtgcagagaa gctgaagaaa cgaatgcagg
     1441 gccccattcc agagcgaatc ctgggcaaga tgactgtggc gattgtgaaa gcactgtact
     1501 atctgaagga gaagcatggc gtcatccatc gcgatgtcaa accctccaac atcctgctag
     1561 atgagcgggg ccagatcaag ctctgtgact ttggcatcag tggccgcctt gttgactcca
     1621 aagccaaaac acggagtgct ggctgtgctg cctatatggc tcccgagcgc atcgaccctc
     1681 cagatcccac caagcctgac tatgacatcc gagctgatgt gtggagcctg ggcatctcac
     1741 tggtggagct ggcaacagga cagttcccct ataagaactg caagacggac tttgaggtcc
     1801 tcaccaaagt cctacaggaa gagcccccac tcctgcctgg tcacatgggc ttctcagggg
     1861 acttccagtc atttgtcaaa gactgcctta ctaaagatca caggaagaga ccaaagtata
     1921 ataagctact tgaacacagc ttcatcaagc actatgagat actcgaggtg gatgtcgcgt
     1981 cctggtttaa ggatgtcatg gcgaagaccg agtccccaag gactagtgga gtcctgagtc
     2041 agcaccatct gcccttcttc agtgggagtc tggaggagtc tcccacttcc ccaccttctc
     2101 ccaagtcctt ccctctgtca ccagccatcc ctcacgaaat tctgagccgt cgtccgagtt
     2161 accgtaaagg atccatgagc agaagcaagc gtgacaacaa tttttatagt gtagagattg
     2221 gagattctac attcacagtc ctgaaacgat atcagaattt aaaacctata ggctcaggag
     2281 ctcaaggaat agtatgcgca gcttatgatg ccattcttga aagaaatgtt gcaatcaaga
     2341 agctaagccg accatttcag aatcagactc atgccaagcg ggcctacaga gagctagttc
     2401 ttatgaaatg tgttaatcac aaaaatataa ttggcctttt gaatgttttc acaccacaga
     2461 aatccctaga agaatttcaa gatgtttaca tagtcatgga gctcatggat gcaaatcttt
     2521 gccaagtgat tcagatggag ctagatcatg aaagaatgtc ctaccttctc tatcagatgc
     2581 tgtgtggaat caagcacctt cattctgctg gaattattca tcgggactta aagcccagta
     2641 atatagtagt aaaatctgat tgcactttga agattcttga cttcggtctg gccaggactg
     2701 caggaacgag ttttatgatg gcgccttttg tcgtcacacg gtattacaga gcacccgagg
     2761 tcatccttgg catgggctac aaggaaaacg tggatttatg gtctgtgggg tgcattatgg
     2821 gagaaatggt ttgccacaaa atcctctttc caggaaggga ctatattgat cagtggaata
     2881 aagttattga acagcttgga acaccatgtc ctgaattcat gaagaaactg caaccaacag
     2941 taaggactta cgttgaaaac agacctaaat atgctggata tagctttgag aaactcttcc
     3001 ctgatgtcct tttcccagct gactcagaac acaacaaact taaagccagt caggcaaggg
     3061 atttgttatc caaaatgctg gtaatagatg catctaaaag gatctctgta gatgaagctc
     3121 tccaacaccc gtacatcaat gtctggtatg atccttctga agcagaagct ccaccaccaa
     3181 agatccctga caagcagtta gatgaaaggg aacacacaat agaagagtgg aaagaattga
     3241 tatataagga agttatggac ttggaggaga gaaccaagaa tggagttata cgggggcagc
     3301 cctctccttt agcacaggtg cagcagtgac tcgagctcaa gcttcaggcc accagtgttg
     3361 ccctcttccc tttttaaaac aaaataccct tgtttgtaaa tccttagacg cttgagaata
     3421 aaacccttcc cttttcttcc gaaaaaaaaa aaaaaaaaac tcgagggggg ggccctattc
     3481 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     3541 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3601 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3661 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3721 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3781 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3841 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3901 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3961 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4021 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4081 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4141 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4201 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4501 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4561 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4621 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4681 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4741 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4801 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4861 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4921 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4981 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5041 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5101 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5161 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5221 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5281 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5341 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5401 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5461 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5521 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5581 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5641 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5701 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5761 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5821 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7801 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7921 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     7981 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     8041 agtgcgcgag ca
//
