LOCUS       sequence-473620-        6560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of DUSP6 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6560)
  AUTHORS   Duan S, Moro L, Qu R, Simoneschi D, Cho H, Jiang S, Zhao H, Chang Q,
            de Stanchina E, Arbini AA, Pagano M
  TITLE     Loss of FBXO31-mediated degradation of DUSP6 dysregulates ERK and 
            PI3K-AKT signaling and promotes prostate tumorigenesis.
  JOURNAL   Cell Rep. 2021 Oct 19;37(3):109870. doi: 
            10.1016/j.celrep.2021.109870.
  PUBMED    34686346
REFERENCE   2  (bases 1 to 6560)
  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: "Cell Rep.";
            date: "2021-10-19"; volume: "37(3):109870. doi"; pages: " 
            10.1016/j.celrep.2021.109870"
COMMENT     Sequence Label: pcDNA HA DUSP6
FEATURES             Location/Qualifiers
     source          1..6560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..58)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     649..668
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    953..1074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(990..1009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1044..1063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1123..1139)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1123..1139)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1136..1158)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1147..1163
                     /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(1171..1177,1178..1195,1196..1201))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1216..1237
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1354..1371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1525..2113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1605..1624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2284..3075,3076..3144))
                     /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     2907..2926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3145..3249)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3324..3343)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3515..3894
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3895..4098
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4049..4069
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4143..4162
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4143..4161
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4196..4222
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(5393..5411)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5394..5411)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5431..5448)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5437..5661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5707..6135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5794..5813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6004..6025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6144..6164)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6149..6478
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6329..6464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6391..6410
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(6545..6560,1..779)
                     /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"
ORIGIN
        1 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
       61 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
      121 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
      181 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
      241 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
      301 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
      361 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
      421 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
      481 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
      541 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
      601 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
      661 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
      721 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
      781 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
      841 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
      901 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
      961 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1021 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1081 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     1141 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1201 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1261 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1321 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1381 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1441 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1501 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1561 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1621 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1681 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1741 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1801 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1861 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1921 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1981 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2041 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2101 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2161 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2221 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2281 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2341 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2401 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2461 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2521 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2581 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2641 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2701 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2761 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2821 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2881 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2941 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3001 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3061 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3121 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3181 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3241 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     3301 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3361 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     3421 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     3481 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     3541 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     3601 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     3661 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     3721 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     3781 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     3841 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     3901 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     3961 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     4021 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     4081 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     4141 attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ccatgtaccc
     4201 atacgatgtt ccagattacg ctatagatac gctcagaccc gtgcccttcg cgtcggaaat
     4261 ggcgatcagc aagacggtgg cgtggctcaa cgagcagctg gagctgggca acgagcggct
     4321 gctgctgatg gactgccggc cgcaggagct atacgagtcg tcgcacatcg agtcggccat
     4381 caacgtggcc atcccgggca tcatgctgcg gcgcctgcag aagggtaacc tgccggtgcg
     4441 cgcgctcttc acgcgcggcg aggaccggga ccgcttcacc cggcgctgtg gcaccgacac
     4501 agtggtgctc tacgacgaga gcagcagcga ctggaacgag aatacgggcg gcgagtcgtt
     4561 gctcgggctg ctgctcaaga agctcaagga cgagggctgc cgggcgttct acctggaagg
     4621 tggcttcagt aagttccaag ccgagttctc cctgcattgc gagaccaatc tagacggctc
     4681 gtgtagcagc agctcgccgc cgttgccagt gctggggctc gggggcctgc ggatcagctc
     4741 tgactcttcc tcggacatcg agtctgacct tgaccgagac cccaatagtg caacagactc
     4801 ggatggtagt ccgctgtcca acagccagcc ttccttccca gtggagatct tgcccttcct
     4861 ctacttgggc tgtgccaaag actccaccaa cttggacgtg ttggaggaat tcggcatcaa
     4921 gtacatcttg aacgtcaccc ccaatttgcc gaatctcttt gagaacgcag gagagtttaa
     4981 atacaagcaa atccccatct cggatcactg gagccaaaac ctgtcccagt ttttccctga
     5041 ggccatttct ttcatagatg aagcccgggg caagaactgt ggtgtcttgg tacattgctt
     5101 ggctggcatt agccgctcag tcactgtgac tgtggcttac cttatgcaga agctcaatct
     5161 gtcgatgaac gatgcctatg acattgtcaa aatgaaaaaa tccaacatat cccctaactt
     5221 caacttcatg ggtcagctgc tggacttcga gaggacgctg ggactcagca gcccatgtga
     5281 caacagggtt ccagcacagc agctgtattt taccacccct tccaaccaga atgtatacca
     5341 ggtggactct ctgcaatcta cgtgactcga gcatgcatct agagggccct attctatagt
     5401 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     5461 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5521 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5581 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     5641 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     5701 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6121 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     6181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     6241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6301 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     6421 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     6481 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     6541 tcgcatgatt gaacaagatg
//
