LOCUS       sequence-192052-        6241 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 6241)
  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 6241)
  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: pRc/RSV Flag MKK3(glu)
FEATURES             Location/Qualifiers
     source          1..6241
                     /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"
     promoter        342..603
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     CDS             627..650
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1740..1757)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1746..1970
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2016..2529
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2103..2122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2313..2334
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2552..2572)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2557..2886
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2737..2872
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2953..3747
                     /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(3007..3026)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3617..3636
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3921..4055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3958..3977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4012..4031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4086..4102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4086..4102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4099..4121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4110..4126
                     /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(4134..4140,4141..4158,4159..4164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4179..4200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4317..4334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4488..5076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4568..4587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5247..6038,6039..6107))
                     /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     5870..5889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6108..6212)
                     /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 cgcgtatctg
      241 aggggactag ggtgtgttta ggcgaaaagc ggggcttcgg ttgtacgcgg ttaggagtcc
      301 cctcaggata tagtagtttc gcttttgcat agggaggggg aaatgtagtc ttatgcaata
      361 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
      421 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
      481 caacagacgg gtctgacatg gattggacga accactgaat tccgcattgc agagatattg
      541 tatttaagtg cctagctcga tacaataaac gccatttgac cattcaccac attggtgtgc
      601 acctccaagc ttcggcagcc aacatggact ataaggacga tgacgacaaa tccaagccac
      661 ccgcacccaa ccccacaccc ccccggaacc tggactcccg gaccttcatc accattggag
      721 acagaaactt tgaggtggag gctgatgact tggtgaccat ctcagaactg ggccgtggag
      781 cctatggggt ggtagagaag gtgcggcacg cccagagcgg caccatcatg gccgtgaagc
      841 ggatccgggc caccgtgaac tcacaggagc agaagcggct gctcatggac ctggacatca
      901 acatgcgcac ggtcgactgt ttctacactg tcaccttcta cggggcacta ttcagagagg
      961 gagacgtgtg gatctgcatg gagctcatgg acacatcctt ggacaagttc taccggaagg
     1021 tgctggataa aaacatgaca attccagagg acatccttgg ggagattgct gtgtctatcg
     1081 tgcgggccct ggagcatctg cacagcaagc tgtcggtgat ccacagagat gtgaagccct
     1141 ccaatgtcct tatcaacaag gagggccatg tgaagatgtg tgactttggc atcagtggct
     1201 acttggtgga cgaggtggcc aaggagatgg atgccggctg caagccctac atggcccctg
     1261 agaggatcaa cccagagctg aaccagaagg gctacaatgt caagtccgac gtctggagcc
     1321 tgggcatcac catgattgag atggccatcc tgcggttccc ttacgagtcc tgggggaccc
     1381 cgttccagca gctgaagcag gtggtggagg agccgtcccc ccagctccca gccgaccgtt
     1441 tctcccccga gtttgtggac ttcactgctc agtgcctgag gaagaacccc gcagagcgta
     1501 tgagctacct ggagctgatg gagcacccct tcttcacctt gcacaaaacc aagaagacgg
     1561 acattgctgc cttcgtgaag aagatcctgg gagaagactc ataggggctg ggcctcggac
     1621 aagccgaatt ccagcacact ggcggccgtt actagtaacg gccgccagtg tgctggaatt
     1681 ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagctc gctgatcagc
     1741 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1801 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1861 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1921 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     1981 ggaaagaacc agctggggct cgagggggga tccccacgcg ccctgtagcg gcgcattaag
     2041 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2101 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2161 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2221 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2281 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2341 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2401 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2461 gcttacaatt taaatatttg cttatacaat cttcctgttt ttggggcttt tctgattatc
     2521 aaccggggtg ggtaccgagc tcgaattctg tggaatgtgt gtcagttagg gtgtggaaag
     2581 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2641 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2701 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2761 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2821 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2881 tgcaaaaagc tcccgggagc ttggatatcc attttcggat ctgatcaaga gacaggatga
     2941 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3001 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3061 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3121 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3181 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3241 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3301 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3361 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3421 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3481 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3541 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3601 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3661 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3721 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3781 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3841 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3901 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     3961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4021 ccaaactcat caatgtatct tatcatgtct ggatccgtcg acctcgagag cttggcgtaa
     4081 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4141 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4201 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4261 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4321 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4381 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4441 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4501 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4561 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4621 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4681 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4741 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4801 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4861 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4921 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4981 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5041 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5101 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5161 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5221 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5281 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5341 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5401 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5461 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5521 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5581 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5641 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5701 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5761 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5821 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5881 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5941 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6001 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6061 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6181 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6241 g
//
