LOCUS       sequence-8422-ex        7151 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 7151)
  AUTHORS   Woodgett JR
  TITLE     Molecular cloning and expression of glycogen synthase 
            kinase-3/factor A.
  JOURNAL   EMBO J. 1990 Aug . 9(8):2431-8.
  PUBMED    2164470
REFERENCE   2  (bases 1 to 7151)
  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: "EMBO J. 
            1990 Aug . 9(8):2431-8."
COMMENT     Sequence Label: GSK3 alpha pMT2
FEATURES             Location/Qualifiers
     source          1..7151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(7..336)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      21..156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     321..341
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          805..1034
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1017..1036
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3351..3914
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    4102..4236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4193..4212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4763..4780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4934..5522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5014..5033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5693..6484,6485..6553))
                     /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     6316..6335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6554..6658)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6726..6744
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6782..6804)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6904..6923
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7117..7139
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7131..7148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7132..7148
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
       61 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
      121 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
      181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      241 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      301 ggagcctggg gactttccac accctaactg acacacattc cacaggatcc ggtcgcgcga
      361 atttcgagcg gtgttccgcg gtcctcctcg tatagaaact cggaccactc tgagacgaag
      421 gctcgcgtcc aggccagcac gaaggaggct aagtgggagg ggtagcggtc gttgtccact
      481 agggggtcca ctcgctccag ggtgtgaaga cacatgtcgc cctcttcggc atcaaggaag
      541 gtgattggtt tataggtgta ggccacgtga ccgggtgttc ctgaaggggg gctataaaag
      601 ggggtggggg cgcgttcgtc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt
      661 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc
      721 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga
      781 tcggaaaacc tctcgactgt tggggtgagt actccctctc aaaagcgggc atgacttctg
      841 cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgcggtga
      901 tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg ttgtcaagct
      961 tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca tccactttgc
     1021 ctttctctcc acaggtgtcc actcccaggt ccaactgcag gcgagcctga attcgccaga
     1081 gcggcgcggc ctggaagagg ccagggcccg ggggaggcga cggcagcggc ggcggctggg
     1141 gcagcccggg cagcccgagc cccgcagcct gggcctgtgc tcggcgccat gagcggcggc
     1201 gggccttcgg gaggcggccc tgggggctcg ggcagggcgc ggactagctc gttcgcggag
     1261 cccggcggcg gaggcggagg aggcggcggc ggccccggag gctcggcctc cggcccaggc
     1321 ggcaccggcg gcggaaaggc atctgtcggg gccatgggtg ggggcgtcgg ggcctcgagc
     1381 tccgggggtg gacccggcgg cagcggcgga ggaggcagcg gaggccccgg cgcaggcact
     1441 agcttcccgc cgcccggggt gaagctgggc cgtgacagcg ggaaggtgac cacagtcgta
     1501 gccactctag gccaaggccc agagcgctcc caagaagtgg cttacacgga catcaaagtg
     1561 attggcaatg gctcatttgg ggtcgtgtac caggcacggc tggcagagac cagggaacta
     1621 gtcgccatca agaaggttct ccaggacaag aggttcaaga accgagagct gcagatcatg
     1681 cgtaagctgg accactgcaa tattgtgagg ctgagatact ttttctactc cagtggcgag
     1741 aagaaagacg agctttacct aaatctggtg ctggaatatg tgcccgagac agtgtaccgg
     1801 gtggcccgcc acttcaccaa ggccaagttg accatcccta tcctctatgt caaggtgtac
     1861 atgtaccagc tcttccgcag cttggcctac atccactccc agggcgtgtg tcaccgcgac
     1921 atcaagcccc agaacctgct ggtggaccct gacactgctg tcctcaagct ctgcgatttt
     1981 ggcagtgcaa agcagttggt ccgaggggag cccaatgtct cctacatctg ttctcgctac
     2041 taccgggccc cagagctcat ctttggagcc actgattaca cctcatccat cgatgtttgg
     2101 tcagctggct gtgtactggc agagctcctc ttgggccagc ccatcttccc tggggacagt
     2161 ggggtggacc agctggtgga gatcatcaag gtgctgggaa caccaacccg ggaacaaatc
     2221 cgagagatga accccaacta cacggagttc aagttccctc agattaaagc tcacccctgg
     2281 acaaaggtgt tcaaatctcg aacgccgcca gaggccatcg cgctctgctc tagcctgctg
     2341 gagtacaccc catcctcaag gctctcccca ctagaggcct gtgcgcacag cttctttgat
     2401 gaactgcgat gtctgggaac ccagctgcct aacaaccgcc cacttccccc tctcttcaac
     2461 ttcagtgctg gtgaactctc catccaaccg tctctcaacg ccattctcat ccctcctcac
     2521 ttgaggtccc ccagcggcac taccaccctc accccgtcct cacaagcttt aactgagact
     2581 ccgaccagct cagactggca gtcgaccgat gccacaccta ccctcactaa ctcctcctga
     2641 gggccccacc aagcaccctt ccacttccat ctgggagccc caagagggcg tgggaagggg
     2701 ggccatagcc catcaagctc ctgccctggc tgggccccta gactagaggg cagaggtaaa
     2761 tgagtccctg tccccacctc cagtccctcc ctcaccagcc tcacccctgt ggtgggcttt
     2821 ttaagaggat tttaactggt tgtggggagg gaagagaagg acagggtgtt ggggggatga
     2881 ggacctccta cccccttggc cccctcccct cccccagacc tccacctcct ccagaccccc
     2941 tcccctcctg tgtcccttgt aaatagaacc agcccagccc gtctcctctt cccttccctg
     3001 gcccccgggt gtaaatagat tgttataatt tttttcttaa agaaaacgtc gattcgcacc
     3061 gtccaacctg ccccgcccct cctacagctg taactcccct cctgtcctct gcccccaagg
     3121 tctactccct cctcacccca ccctggaggg ccaggggagt ggagagagct cctgatgtct
     3181 tagtttccac agtaaggttt gcctgtgtac agacctccgt tcaataaatt attggcatga
     3241 aaagaattcg gggggggggg ggggggggac agctcagggc tgcgatttcg cgccaaactt
     3301 gacggcaatc ctagcgtgaa ggctggtagg attttatccc cgctgccatc atggttcgac
     3361 cattgaactg catcgtcgcc gtgtcccaaa atatggggat tggcaagaac ggagacctac
     3421 cctggcctcc gctcaggaac gagttcaagt acttccaaag aatgaccaca acctcttcag
     3481 tggaaggtaa acagaatctg gtgattatgg gtaggaaaac ctggttctcc attcctgaga
     3541 agaatcgacc tttaaaggac agaattaata tagttctcag tagagaactc aaagaaccac
     3601 cacgaggagc tcattttctt gccaaaagtt tggatgatgc cttaagactt attgaacaac
     3661 cggaattggc aagtaaagta gacatggttt ggatagtcgg aggcagttct gtttaccagg
     3721 aagccatgaa tcaaccaggc cacctcagac tctttgtgac aaggatcatg caggaatttg
     3781 aaagtgacac gtttttccca gaaattgatt tggggaaata taaacttctc ccagaatacc
     3841 caggcgtcct ctctgaggtc caggaggaaa aaggcatcaa gtataagttt gaagtctacg
     3901 agaagaaaga ctaacaggaa gatgctttca agttctctgc tcccctccta aagctatgca
     3961 ttttttataa gaccatggga cttttgctgg ctttagatca taatcagcca taccacattt
     4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4201 tccaaactca tcaatgtatc ttatcatgtc tggatccccg gccaacggtc tggtgacccg
     4261 gctgcgagag ctcggtgtac ctgagacgcg agtaagccct tgagtcaaag acgtagtcgt
     4321 tgcaagtccg caccaggtac tgatatccca ccaaaaagtg cggcggcggc tggcggtaga
     4381 ggggccagcg tagggtggcc ggggctccgg gggcgaggtc ttccaacata aggcgatgat
     4441 catcgatgct agaccgtgca aaaggagagc ctgtaagcgg gcactcttcc gtggtctggt
     4501 ggataaattc gcaagggtat catggcggac gaccggggtt cgaaccccgg atccggccgt
     4561 ccgccgtgat ccatccggtt accgcccgcg tgtcgaaccc aggtgtgcga cgtcagacaa
     4621 cgggggagcg ctccttttgg cttccttcca ggcgcggcgg ctgctgcgct agcttttttg
     4681 gcgagctcga attaattctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4741 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4801 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4861 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5101 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     5161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5341 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6661 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6721 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6781 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6841 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6901 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     6961 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     7021 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     7081 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     7141 cggccagtgc c
//
