LOCUS       sequence-352171-        6794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human Gsk3beta with S9A (constitutively active) mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6794)
  AUTHORS   Lang UE, Kocabayoglu P, Cheng GZ, Ghiassi-Nejad Z, Munoz U, Vetter 
            D, Eckstein DA, Hannivoort RA, Walsh MJ, Friedman SL
  TITLE     GSK3beta phosphorylation of the KLF6 tumor suppressor promotes its 
            transactivation of p21.
  JOURNAL   Oncogene. 2012 Oct 22. doi: 10.1038/onc.2012.457.
  PUBMED    23085750
REFERENCE   2  (bases 1 to 6794)
  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: "Oncogene. 
            2012 Oct 22. doi: 10.1038/onc.2012.457."
COMMENT     Sequence Label: GSK3betaS9A  (1016)
FEATURES             Location/Qualifiers
     source          1..6794
                     /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             1568..1579
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2014..2040
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2020..2046
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(2113..2131)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2114..2131)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2151..2168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2157..2381
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2427..2855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2514..2533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2724..2745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2864..2884)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2869..3198
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3049..3184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3111..3130
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3265..4059
                     /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(3319..3338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3929..3948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4233..4354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4270..4289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4324..4343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4403..4419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4403..4419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4416..4438)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4427..4443
                     /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(4451..4457,4458..4475,4476..4481))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4496..4517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4634..4651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4805..5393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4885..4904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5564..6355,6356..6424))
                     /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     6187..6206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6425..6529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6604..6623)
                     /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 tcggatccac tagtaacggc cgccagtgtg ctggaattcc ggaggaagga
      721 aggaaaaggt gattcgcgaa gagagtgatc atgtcagggc ggcccagaac caccgccttt
      781 gcggagagct gcaagccggt gcagcagcct tcagcttttg gcagcatgaa agttagcaga
      841 gacaaggacg gcagcaaggt gacaacagtg gtggcaactc ctgggcaggg tccagacagg
      901 ccacaagaag tcagctatac agacactaaa gtgattggaa atggatcatt tggtgtggta
      961 tatcaagcca aactttgtga ttcaggagaa ctggtcgcca tcaagaaagt attgcaggac
     1021 aagagattta agaatcgaga gctccagatc atgagaaagc tagatcactg taacatagtc
     1081 cgattgcgtt atttcttcta ctccagtggt gagaagaaag atgaggtcta tcttaatctg
     1141 gtgctggact atgttccgga aacagtatac agagttgcca gacactatag tcgagccaaa
     1201 cagacgctcc ctgtgattta tgtcaagttg tatatgtatc agctgttccg aagtttagcc
     1261 tatatccatt cctttggaat ctgccatcgg gatattaaac cgcagaacct cttgttggat
     1321 cctgatactg ctgtattaaa actctgtgac tttggaagtg caaagcagct ggtccgagga
     1381 gaacccaatg tttcgtatat ctgttctcgg tactataggg caccagagtt gatctttgga
     1441 gccactgatt atacctctag tatagatgta tggtctgctg gctgtgtgtt ggctgagctg
     1501 ttactaggac aaccaatatt tccaggggat agtggtgtgg atcagttggt agaaataatc
     1561 aaggtcctgg gaactccaac aagggagcaa atcagagaaa tgaacccaaa ctacacagaa
     1621 tttaaattcc ctcaaattaa ggcacatcct tggactaagg tcttccgacc ccgaactcca
     1681 ccggaggcaa ttgcactgtg tagccgtctg ctggagtata caccaactgc ccgactaaca
     1741 ccactggaag cttgtgcaca ttcatttttt gatgaattac gggacccaaa tgtcaaacta
     1801 ccaaatgggc gagacacacc tgcactcttc aacttcacca ctcaagaact gtcaagtaat
     1861 ccacctctgg ctaccatcct tattcctcct catgctcgga ttcaagcagc tgcttcaacc
     1921 cccacaaatg ccacagcagc gtcagatgct aatactggag accgtggaca gaccaataat
     1981 gctgcttctg catcagcttc caactccact agttacccat acgatgttcc agattacgca
     2041 agcttgggtg gtcccaacta gtacagtccc gacggatcct ctagagtcga gcatgcatct
     2101 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     2161 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2221 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2281 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2341 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2401 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2461 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2521 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2581 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2641 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2701 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2761 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2821 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2881 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2941 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3001 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3061 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3121 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3181 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3241 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3301 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3361 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3421 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3481 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3541 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3601 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3661 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3721 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3781 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3841 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3901 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3961 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4021 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4081 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4141 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4201 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4261 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4321 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4381 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4441 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4501 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4561 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4621 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4681 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4741 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4981 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5221 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5281 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5341 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5401 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5461 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5521 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5581 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5641 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5701 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5761 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5821 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5881 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5941 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6001 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6061 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6121 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6181 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6241 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6301 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6361 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6421 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6481 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6541 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     6601 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     6661 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     6721 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     6781 cagatatacg cgtt
//
