LOCUS       sequence-222741-        6589 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 6589)
  AUTHORS   Raingeaud J, Whitmarsh AJ, Barrett T, Derijard B, Davis RJ
  TITLE     MKK3- and MKK6-regulated gene expression is mediated by the p38 
            mitogen-activated protein kinase signal transduction pathway.
  JOURNAL   Mol Cell Biol. 1996 Mar . 16(3):1247-55.
  PUBMED    8622669
REFERENCE   2  (bases 1 to 6589)
  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. 1996 Mar . 16(3):1247-55."
COMMENT     Sequence Label: pCDNA3-Flag MKK6
FEATURES             Location/Qualifiers
     source          1..6589
                     /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             911..934
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2144..2162)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2145..2162)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2182..2199)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2188..2412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2458..2886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2545..2564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2755..2776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2895..2915)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2900..3229
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3080..3215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3142..3161
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3296..4090
                     /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(3350..3369)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3960..3979
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4264..4385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4301..4320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4355..4374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4434..4450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4434..4450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4447..4469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4458..4474
                     /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(4482..4488,4489..4506,4507..4512))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4527..4548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4665..4682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4836..5424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4916..4935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5595..6386,6387..6455))
                     /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     6218..6237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6456..6560)
                     /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 aagcttcggc
      901 agccaacatg gactataagg acgatgatga caaatctcag tcgaaaggca agaagcgaaa
      961 ccctggcctt aaaattccaa aagaagcatt tgaacaacct cagaccagtt ccacaccacc
     1021 tagagattta gactccaagg cttgcatttc tattggaaat cagaactttg aggtgaaggc
     1081 agatgacctg gagcctataa tggaactggg acgaggtgcg tacggggtgg tggagaagat
     1141 gcggcacgtg cccagcgggc agatcatggc agtgaagcgg atccgagcca cagtaaatag
     1201 ccaggaacag aaacggctac tgatggattt ggatatttcc atgaggacgg tggactgtcc
     1261 attcactgtc accttttatg gcgcactgtt tcgggagggt gatgtgtgga tctgcatgga
     1321 gctcatggat acatcactag ataaattcta caaacaagtt attgataaag gccagacaat
     1381 tccagaggac atcttaggga aaatagcagt ttctattgta aaagcattag aacatttaca
     1441 tagtaagctg tctgtcattc acagagacgt caagccttct aatgtactca tcaatgctct
     1501 cggtcaagtg aagatgtgcg attttggaat cagtggctac ttggtggact ctgttgctaa
     1561 aacaattgat gcaggttgca aaccatacat ggcccctgaa agaataaacc cagagctcaa
     1621 ccagaaggga tacagtgtga agtctgacat ttggagtctg ggcatcacga tgattgagtt
     1681 ggccatcctt cgatttccct atgattcatg gggaactcca tttcagcagc tcaaacaggt
     1741 ggtagaggag ccatcgccac aactcccagc agacaagttc tctgcagagt ttgttgactt
     1801 tacctcacag tgcttaaaga agaattccaa agaacggcct acatacccag agctaatgca
     1861 acatccattt ttcaccctac atgaatccaa aggaacagat gtggcatctt ttgtaaaact
     1921 gattcttgga gactaaaaag cagtggactt aatcggttga ccctactgtg gattggtggg
     1981 tttcggggtg aagcaagttc actacagcat caatagaaag tcatctttga gataatttaa
     2041 ccctgcctct cagagggttt tctctcccaa ttttcttttt actccccctc ttaagggggc
     2101 cttggaatct atagtataga atgaactgtc tagagggccc tattctatag tgtcacctaa
     2161 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2221 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2281 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2341 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2401 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2461 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2521 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2581 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2641 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2701 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2761 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2821 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2881 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3001 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3061 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3121 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3241 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3301 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3361 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3421 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3481 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3541 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3601 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3661 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3721 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3781 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3841 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3901 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3961 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4021 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4081 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4141 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4201 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4261 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4321 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4381 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4441 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4501 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4561 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4621 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4681 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4741 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4801 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4861 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4921 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4981 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5041 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5101 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5161 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5221 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5281 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5341 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5401 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5461 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5521 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5581 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5641 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5701 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5761 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5821 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5881 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5941 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6001 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6061 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6121 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6181 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6241 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6301 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6361 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6421 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
