LOCUS       sequence-341893-        6582 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 6582)
  AUTHORS   Enslen H, Raingeaud J, Davis RJ
  TITLE     Selective activation of p38 mitogen-activated protein (MAP) kinase 
            isoforms by the MAP kinase kinases MKK3 and MKK6.
  JOURNAL   J Biol Chem. 1998 Jan 16. 273(3):1741-8.
  PUBMED    9430721
REFERENCE   2  (bases 1 to 6582)
  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: "J Biol 
            Chem. 1998 Jan 16. 273(3):1741-8."
COMMENT     Sequence Label: pCDNA3 Flag p38 beta2
FEATURES             Location/Qualifiers
     source          1..6582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             685..708
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1901..1919)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1902..1919)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1939..1956)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1945..2169
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2215..2643
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2302..2321)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2512..2533
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2652..2672)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2657..2986
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2837..2972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2899..2918
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3053..3847
                     /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(3107..3126)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3717..3736
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4021..4142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4058..4077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4112..4131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4191..4207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4191..4207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4204..4226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4215..4231
                     /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(4239..4245,4246..4263,4264..4269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4284..4305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4422..4439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4593..5181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4673..4692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5352..6143,6144..6212))
                     /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     5975..5994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6213..6317)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6392..6411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatctaa gatggactac aaggacgatg acgataagtc gggccctcgc
      721 gccggcttct accggcagga gctgaacaag accgtgtggg aggtgccgct gcggctgcag
      781 gggctgcgcc cggtgggctc cggcgcctac ggctccgtct gttcggccta cgacgcccgg
      841 ctgcgccaga aggtggcggt gaagaagctg tcgcgcccct tccagtcgct gatccacgcg
      901 cgcagaacgt accgggagct gcggctgctc aagcacctga agcacgagaa cgtcatcggg
      961 cttctggacg tcttcacgcc ggccacgtcc atcgaggact tcagcgaagt gtacttggta
     1021 accaccctga tgggcgccga cctgaacaac atcgtcaagt gccaggcgct gagcgacgag
     1081 cacgttcaat tcctggttta ccagctgctg cgcgggctga agtacatcca ctcggccggg
     1141 atcatccacc gggacctgaa gcccagcaac gtggctgtga acgaggactg tgagctcagg
     1201 atcctggatt tcgggctggc gcgccaggcg gacgaggaga tgaccggcta tgtggccacg
     1261 cgctggtacc gggcacctga gatcatgctc aactggttgc attacaacca aacagtggat
     1321 atctggtccg taggctgcat catggctgag ctgctccagg gcaaggccct cttcccggga
     1381 agcgactaca ttgaccagct gaagcgcatc atggaagtgg tgggcacacc cagccctgag
     1441 gttctggcaa aaatctcctc ggaacacgcc cggacatata tccagtccct gccccccatg
     1501 ccccagaagg acctgagcag catcttccgt ggagccaacc ccctggccat agacctcctt
     1561 ggaaggatgc tggtgctgga cagtgaccag agggtcagtg cagctgaggc actggcccac
     1621 gcctacttca gccagtacca cgaccccgag gatgagccag aggccgagcc atatgatgag
     1681 agcgttgagg ccaaggagcg cacgctggag gagtggaagg agctcactta ccaggaagtc
     1741 cttagcttca agcccccaga gccaccgaag ccacctggca gcctggagat tgagcagtga
     1801 ggtgctgccc agcagcccct gagagcctgt ggaggggctt gggcctgcac ccttccacag
     1861 ctggcctggt ttcctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     1921 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1981 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2041 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2101 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2161 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2221 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2281 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2341 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2401 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2461 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2521 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2581 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2641 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2701 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2761 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2821 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2881 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     2941 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3001 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3061 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3121 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3181 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3241 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3301 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3361 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3421 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3481 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3541 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3601 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3661 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3721 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3781 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3841 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     3901 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3961 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4021 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4081 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4141 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4201 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4261 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4321 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4381 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5341 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5401 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5461 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5521 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5581 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5641 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5701 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5761 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5821 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5881 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5941 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6001 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6061 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6121 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6181 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6241 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6301 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     6361 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6421 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     6481 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     6541 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
