LOCUS       sequence-146842-        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(apf)
FEATURES             Location/Qualifiers
     source          1..7211
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2764..2782)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2765..2782)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2802..2819)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2808..3032
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3078..3506
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3165..3184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3375..3396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3515..3535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3520..3849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3700..3835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3762..3781
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3916..4710
                     /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(3970..3989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4580..4599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4884..5005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4921..4940)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4975..4994
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5054..5070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5054..5070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5067..5089)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5078..5094
                     /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(5102..5108,5109..5126,5127..5132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5147..5168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5285..5302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5456..6044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5536..5555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6215..7006,7007..7075))
                     /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     6838..6857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7076..7180)
                     /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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctttcggca
      901 gccaacatgg actataagga cgatgatgac aaaagcgaca gtaaatgtga cagtcagttt
      961 tatagtgtgc aagtggcaga ctcaaccttc actgtcctaa aacgttacca gcagctgaaa
     1021 ccaattggct ctggggccca agggattgtt tgtgctgcat ttgatacagt tcttgggata
     1081 agtgttgcag tcaagaaact aagccgtcct tttcagaacc aaactcatgc aaagagagct
     1141 tatcgtgaac ttgtcctctt aaaatgtgtc aatcataaaa atataattag tttgttaaat
     1201 gtgtttacac cacaaaaaac tctagaagaa tttcaagatg tgtatttggt tatggaatta
     1261 atggatgcta acttatgtca ggttattcac atggagctgg atcatgaaag aatgtcctac
     1321 cttctttacc agatgctttg tggtattaaa catctgcatt cagctggtat aattcataga
     1381 gatttgaagc ctagcaacat tgttgtgaaa tcagactgca ccctgaagat ccttgacttt
     1441 ggcctggccc ggacagcgtg cactaacttc atgatggccc ctttcgtggt gacacggtac
     1501 taccgggcgc ccgaagtcat cctgggtatg ggctacaaag agaacgttga tatctggtca
     1561 gtgggttgca tcatgggaga gctggtgaaa ggttgtgtga tattccaagg cactgaccat
     1621 attgatcagt ggaataaagt tattgagcag ctgggaacac catcagcaga gttcatgaag
     1681 aaacttcagc caactgtgag gaattatgtc gaaaacagac caaagtatcc tggaatcaaa
     1741 tttgaagaac tctttccaga ttggatattc ccatcagaat ctgagcgaga caaaataaaa
     1801 acaagtcaag ccagagatct gttatcaaaa atgttagtga ttgatcctga caagcggatc
     1861 tctgtagacg aagctctgcg tcacccatac atcactgttt ggtatgaccc cgccgaagca
     1921 gaagccccac cacctcaaat ttatgatgcc cagttggaag aaagagaaca tgcaattgaa
     1981 gaatggaaag agctaattta caaagaagtc atggattggg aagaaagaag caagaatggt
     2041 gttgtaaaag atcagccttc agatgcagca gtaagtagca acgccactcc ttctcagtct
     2101 tcatcgatca atgacatttc atccatgtcc actgagcaga cgctggcctc agacacagac
     2161 agcagtcttg atgcctcgac gggacccctt gaaggctgtc gatgataggt tagaaatagc
     2221 aaacctgtca gcattgaagg aactctcacc tccgtgggcc tgaaatgctt gggagttgat
     2281 ggaaccaaat agaaaaactc catgttctgc atgtaagaaa cacaatgcct tgccctattc
     2341 agacctgata ggattgcctg cttagatgat aaaatgaggc agaatatgtc tgaagaaaaa
     2401 aattgcaagc cacacttcta gagattttgt tcaagatcat ttcaggtgag cagttagagt
     2461 aggtgaattt gtttcaaatt gtactagtga cagtttctca tcatctgtaa ctgttgagat
     2521 gtatgtgcat gtgaccacaa atgcttgctt ggacttgccc atctagcact ttggaaatca
     2581 gtatttaaat gccaaataat cttccaggta gtgctgcttc tgaagttatc tcttaatcct
     2641 cttaagtaat ttgggaattc ctgcagcccg ggggatccac tagtaacggc cgccagtgtg
     2701 ctggaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     2761 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     2821 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2881 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2941 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3001 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3061 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3121 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3181 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3241 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3301 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3361 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3421 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3481 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3541 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3601 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3661 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3721 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3781 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3841 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3901 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3961 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4021 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4081 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4141 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4201 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4261 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4321 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4381 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4441 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4501 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4561 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4621 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4681 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4741 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4801 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4861 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4921 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4981 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5041 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5101 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5161 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5221 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5281 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5341 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5401 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5461 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5521 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5581 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5641 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5701 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5761 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5821 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5881 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5941 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6001 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6061 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6121 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6181 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6241 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6301 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6361 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6421 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6481 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6541 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6601 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6661 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6721 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6781 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6841 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6901 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6961 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7021 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7081 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7141 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7201 cacctgacgt c
//
