LOCUS       sequence-228211-        7211 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 7211)
  AUTHORS   Gupta S, Barrett T, Whitmarsh AJ, Cavanagh J, Sluss HK, Derijard B, 
            Davis RJ
  TITLE     Selective interaction of JNK protein kinase isoforms with 
            transcription factors.
  JOURNAL   EMBO J. 1996 Jun 3. 15(11):2760-70.
  PUBMED    8654373
REFERENCE   2  (bases 1 to 7211)
  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: "EMBO J. 
            1996 Jun 3. 15(11):2760-70."
COMMENT     Sequence Label: pCDNA3 Flag Jnk2a2
FEATURES             Location/Qualifiers
     source          1..7211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             912..935
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2766..2784)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2767..2784)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2804..2821)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2810..3034
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3080..3508
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3167..3186)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3377..3398
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3517..3537)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3522..3851
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3702..3837
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3764..3783
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3918..4712
                     /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(3972..3991)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4582..4601
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4886..5007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4923..4942)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4977..4996
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5056..5072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5056..5072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5069..5091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5080..5096
                     /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(5104..5110,5111..5128,5129..5134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5149..5170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5287..5304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5458..6046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5538..5557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6217..7008,7009..7077))
                     /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     6840..6859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7078..7182)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctttcgg
      901 cagccaacat ggactataag gacgatgatg acaaaagcga cagtaaatgt gacagtcagt
      961 tttatagtgt gcaagtggca gactcaacct tcactgtcct aaaacgttac cagcagctga
     1021 aaccaattgg ctctggggcc caagggattg tttgtgctgc atttgataca gttcttggga
     1081 taagtgttgc agtcaagaaa ctaagccgtc cttttcagaa ccaaactcat gcaaaaagag
     1141 cttatcgtga acttgtcctc ttaaaatgtg tcaatcataa aaatataatt agtttgttaa
     1201 atgtgtttac accacaaaaa actctagaag aatttcaaga tgtgtatttg gttatggaat
     1261 taatggatgc taacttatgt caggttattc acatggagct ggatcatgaa agaatgtcct
     1321 accttcttta ccagatgctt tgtggtatta aacatctgca ttcagctggt ataattcata
     1381 gagatttgaa gcctagcaac attgttgtga aatcagactg caccctgaag atccttgact
     1441 ttggcctggc ccggacagcg tgcactaact tcatgatgac cccttacgtg gtgacacggt
     1501 actaccgggc gcccgaagtc atcctgggta tgggctacaa agagaacgtt gatatctggt
     1561 cagtgggttg catcatggga gagctggtga aaggttgtgt gatattccaa ggcactgacc
     1621 atattgatca gtggaataaa gttattgagc agctgggaac accatcagca gagttcatga
     1681 agaaacttca gccaactgtg aggaattatg tcgaaaacag accaaagtat cctggaatca
     1741 aatttgaaga actctttcca gattggatat tcccatcaga atctgagcga gacaaaataa
     1801 aaacaagtca agccagagat ctgttatcaa aaatgttagt gattgatcct gacaagcgga
     1861 tctctgtaga cgaagctctg cgtcacccat acatcactgt ttggtatgac cccgccgaag
     1921 cagaagcccc accacctcaa atttatgatg cccagttgga agaaagagaa catgcaattg
     1981 aagaatggaa agagctaatt tacaaagaag tcatggattg ggaagaaaga agcaagaatg
     2041 gtgttgtaaa agatcagcct tcagatgcag cagtaagtag caacgccact ccttctcagt
     2101 cttcatcgat caatgacatt tcatccatgt ccactgagca gacgctggcc tcagacacag
     2161 acagcagtct tgatgcctcg acgggacccc ttgaaggctg tcgatgatag gttagaaata
     2221 gcaaacctgt cagcattgaa ggaactctca cctccgtggg cctgaaatgc ttgggagttg
     2281 atggaaccaa atagaaaaac tccatgttct gcatgtaaga aacacaatgc cttgccctat
     2341 tcagacctga taggattgcc tgcttagatg ataaaatgag gcagaatatg tctgaagaaa
     2401 aaaattgcaa gccacacttc tagagatttt gttcaagatc atttcaggtg agcagttaga
     2461 gtaggtgaat ttgtttcaaa ttgtactagt gacagtttct catcatctgt aactgttgag
     2521 atgtatgtgc atgtgaccac aaatgcttgc ttggacttgc ccatctagca ctttggaaat
     2581 cagtatttaa atgccaaata atcttccagg tagtgctgct tctgaagtta tctcttaatc
     2641 ctcttaagta atttgggaat tcctgcagcc cgggggatcc actagtaacg gccgccagtg
     2701 tgctggaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc
     2761 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     2821 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2881 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2941 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3001 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3061 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3121 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3181 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3241 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3301 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3361 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3421 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3481 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3541 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3601 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3661 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3721 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3781 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3841 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3901 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3961 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4021 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4081 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4141 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4201 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4261 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4321 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4381 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4441 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4501 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4561 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4621 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4681 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4741 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4801 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4861 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4921 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4981 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5041 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5101 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5161 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5221 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5281 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5341 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5401 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5461 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5521 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5581 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5641 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5701 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5761 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5821 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5881 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5941 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6001 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6061 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6421 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6481 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6541 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6601 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6661 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6721 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6781 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6841 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6901 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6961 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7021 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7081 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7141 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7201 gccacctgac g
//
