LOCUS       sequence-174147-        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
FEATURES             Location/Qualifiers
     source          1..8052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        508..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     662..682
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     756..775
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        756..774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             802..825
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(952..963)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(3498..3516)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3499..3516)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3536..3553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3542..3766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3812..4240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3899..3918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4109..4130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4249..4269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4254..4583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4434..4569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4496..4515
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4650..5444
                     /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(4704..4723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5314..5333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5618..5739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5655..5674)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5709..5728
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5788..5804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5788..5804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5801..5823)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5812..5828
                     /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(5836..5842,5843..5860,5861..5866))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5881..5902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6019..6036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6190..6778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6270..6289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6949..7740,7741..7809))
                     /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     7572..7591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7810..7914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7989..8008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
       61 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      121 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      361 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      421 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      481 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      541 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      601 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      721 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      781 caagcttcgg cagccaacat ggactataag gacgatgatg acaaaatggc ggcgtcctcc
      841 ctggagcaga agctgtcccg cctggaagcc aagctgaagc aggagaaccg tgaggcccgc
      901 aggaggatcg acctcaactt ggatatcagc ccacagcggc ccaggcccac cctgcaactc
      961 ccactggcca acgatggggg cagccgctca ccatcctcag agagctcccc acagcaccct
     1021 acacccccca cccggccccg ccacatgctg gggctcccat caaccttgtt cacaccgcgc
     1081 agtatggaga gcatcgagat tgaccagaag ctgcaggaga tcatgaagca gacagggtac
     1141 ctgactatcg ggggccagcg ttatcaggca gaaatcaatg acttggagaa cttgggtgag
     1201 atgggcagtg gtacctgtgg tcaggtgtgg aagatgcggt tccggaagac aggccacatc
     1261 attgctgtta agcaaatgcg gcgctctggg aacaaggaag agaataagcg cattttgatg
     1321 gacctggatg tagtactcaa gagccatgac tgcccttaca tcgttcagtg ctttggcacc
     1381 ttcatcacca acacagacgt ctttattgcc atggagctca tgggcacatg tgcagagaag
     1441 ctgaagaaac gaatgcaggg ccccattcca gagcgaatcc tgggcaagat gactgtggcg
     1501 attgtgaaag cactgtacta tctgaaggag aagcatggcg tcatccatcg cgatgtcaaa
     1561 ccctccaaca tcctgctaga tgagcggggc cagatcaagc tctgtgactt tggcatcagt
     1621 ggccgccttg ttgactccaa agccaaaaca cggagtgctg gctgtgctgc ctatatggct
     1681 cccgagcgca tcgaccctcc agatcccacc aagcctgact atgacatccg agctgatgtg
     1741 tggagcctgg gcatctcact ggtggagctg gcaacaggac agttccccta taagaactgc
     1801 aagacggact ttgaggtcct caccaaagtc ctacaggaag agcccccact cctgcctggt
     1861 cacatgggct tctcagggga cttccagtca tttgtcaaag actgccttac taaagatcac
     1921 aggaagagac caaagtataa taagctactt gaacacagct tcatcaagca ctatgagata
     1981 ctcgaggtgg atgtcgcgtc ctggtttaag gatgtcatgg cgaagaccga gtccccaagg
     2041 actagtggag tcctgagtca gcaccatctg cccttcttca gtgggagtct ggaggagtct
     2101 cccacttccc caccttctcc caagtccttc cctctgtcac cagccatccc tcacgaaatt
     2161 ctgagccgtc gtccgagtta ccgtaaagga tccatgagca gaagcaagcg tgacaacaat
     2221 ttttatagtg tagagattgg agattctaca ttcacagtcc tgaaacgata tcagaattta
     2281 aaacctatag gctcaggagc tcaaggaata gtatgcgcag cttatgatgc cattcttgaa
     2341 agaaatgttg caatcaagaa gctaagccga ccatttcaga atcagactca tgccaagcgg
     2401 gcctacagag agctagttct tatgaaatgt gttaatcaca aaaatataat tggccttttg
     2461 aatgttttca caccacagaa atccctagaa gaatttcaag atgtttacat agtcatggag
     2521 ctcatggatg caaatctttg ccaagtgatt cagatggagc tagatcatga aagaatgtcc
     2581 taccttctct atcagatgct gtgtggaatc aagcaccttc attctgctgg aattattcat
     2641 cgggacttaa agcccagtaa tatagtagta aaatctgatt gcactttgaa gattcttgac
     2701 ttcggtctgg ccaggactgc aggaacgagt tttatgatga cgccttatgt agtgactcgc
     2761 tactacagag cacccgaggt catccttggc atgggctaca aggaaaacgt ggatttatgg
     2821 tctgtggggt gcattatggg agaaatggtt tgccacaaaa tcctctttcc aggaagggac
     2881 tatattgatc agtggaataa agttattgaa cagcttggaa caccatgtcc tgaattcatg
     2941 aagaaactgc aaccaacagt aaggacttac gttgaaaaca gacctaaata tgctggatat
     3001 agctttgaga aactcttccc tgatgtcctt ttcccagctg actcagaaca caacaaactt
     3061 aaagccagtc aggcaaggga tttgttatcc aaaatgctgg taatagatgc atctaaaagg
     3121 atctctgtag atgaagctct ccaacacccg tacatcaatg tctggtatga tccttctgaa
     3181 gcagaagctc caccaccaaa gatccctgac aagcagttag atgaaaggga acacacaata
     3241 gaagagtgga aagaattgat atataaggaa gttatggact tggaggagag aaccaagaat
     3301 ggagttatac gggggcagcc ctctccttta gcacaggtgc agcagtgact cgagctcaag
     3361 cttcaggcca ccagtgttgc cctcttccct ttttaaaaca aaataccctt gtttgtaaat
     3421 ccttagacgc ttgagaataa aacccttccc ttttcttccg aaaaaaaaaa aaaaaaaact
     3481 cgaggggggg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
     3541 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3601 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3661 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3721 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3781 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4021 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4141 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4261 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4381 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4441 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4501 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4561 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4621 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4681 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4741 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4801 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4861 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4921 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4981 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5041 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5101 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5161 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5221 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5281 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5341 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5401 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5461 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5521 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5581 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5641 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5701 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5761 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5821 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5881 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5941 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7921 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     7981 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     8041 gtgttggagg tc
//
