LOCUS       sequence-315801-        7246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal HA-tagged AXIN1 cDNA expression vector for transfections.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7246)
  AUTHORS   Orozco JM, Krawczyk PA, Scaria SM, Cangelosi AL, Chan SH, Kunchok T,
            Lewis CA, Sabatini DM
  TITLE     Dihydroxyacetone phosphate signals glucose availability to mTORC1.
  JOURNAL   Nat Metab. 2020 Sep;2(9):893-901. doi: 10.1038/s42255-020-0250-5. 
            Epub 2020 Jul 27.
  PUBMED    32719541
REFERENCE   2  (bases 1 to 7246)
  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"; doi: 
            "10.1038/s42255-020-0250-5"; journalName: "Nat Metab"; date: 
            "2020-09"; volume: "2"; issue: "9"; pages: "893-901"
COMMENT     Sequence Label: HA_Axin1 in pRK5
FEATURES             Location/Qualifiers
     source          1..7246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        435..453
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     435..452
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     466..485
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      570..579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             582..608
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3153..3162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3166..3300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3203..3222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3257..3276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3369..3726
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3369..3560
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3392..3412)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3577..3712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3746..3763)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3746..3762)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3755..3777)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3975..4430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4062..4081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4272..4293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4447..4466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4566..4588
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4626..4644)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4712..4816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4817..4885,4886..5677)
                     /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     complement(5035..5054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5848..6436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6337..6356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6590..6607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6724..6745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6760..6765,6766..6783,6784..6790)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6798..6814
                     /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)."
     primer_bind     6803..6825
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6822..6838
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6822..6838
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6867..7246
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
      301 cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc
      361 tataggccca cccccttggc ttcgttagaa cgcggctaca attaatacat aaccttatgt
      421 atcatacaca tacgatttag gtgacactat agaataacat ccactttgcc tttctctcca
      481 caggtgtcca ctcccaggtc caactgcacc tcggttctat cgattgaatt cccgaaccga
      541 cagtcggtct cttcaccaag gccattcgcg ccaccatggg ctacccctat gatgtgcctg
      601 actacgcaga tctcaatggt ggtggtggtg ggtcgacgat gaatatccaa gagcagggtt
      661 tccccttgga cctcggagca agtttcaccg aagatgctcc ccgaccccca gtgcctggtg
      721 aggagggaga actggtgtcc acagacccga ggcccgccag ctacagtttc tgctccggga
      781 aaggtgttgg cattaaaggt gagacttcga cggccactcc gaggcgctcg gatctggacc
      841 tggggtatga gcctgagggc agtgcctccc ccaccccacc atacttgaag tgggctgagt
      901 cactgcattc cctgctggat gaccaagatg ggataagcct gttcaggact ttcctgaagc
      961 aggagggctg tgccgacttg ctggacttct ggtttgcctg cactggcttc aggaagctgg
     1021 agccctgtga ctcgaacgag gagaagaggc tgaagctggc gagagccatc taccgaaagt
     1081 acattcttga taacaatggc atcgtgtccc ggcagaccaa gccagccacc aagagcttca
     1141 taaagggctg catcatgaag cagctgatcg atcctgccat gtttgaccag gcccagaccg
     1201 aaatccaggc cactatggag gaaaacacct atccctcctt ccttaagtct gatatttatt
     1261 tggaatatac gaggacaggc tcggagagcc ccaaagtctg tagtgaccag agctctgggt
     1321 cagggacagg gaagggcata tctggatacc tgccgacctt aaatgaagat gaggaatgga
     1381 agtgtgacca ggacatggac gaggacgatg gcagagacgc tgctcccccc ggaagactcc
     1441 ctcagaagct gctcctggag acagctgccc cgagggtctc ctccagtaga cggtacagcg
     1501 aaggcagaga gttcaggtat ggatcctggc gggagccagt caacccctat tatgtcaatg
     1561 ccggctatgc cctggcccca gccaccagtg ccaacgacag cgagcagcag agcctgtcca
     1621 gcgatgcaga caccctgtcc ctcacggaca gcagcgtgga tgggatcccc ccatacagga
     1681 tccgtaagca gcaccgcagg gagatgcagg agagcgtgca ggtcaatggg cgggtgcccc
     1741 tacctcacat tccccgcacg taccgggtgc cgaaggaggt ccgcgtggag cctcagaagt
     1801 tcgcggagga gctcatccac cgcctggagg ctgtgcagcg cacgcgggag gccgaggaga
     1861 agctggagga gcggctgaag cgcgtgcgca tggaggagga aggtgaggac ggcgatccat
     1921 catcagggcc cccagggccg tgtcacaagc tgcctcccgc ccccgcttgg caccacttcc
     1981 cgccccgctg tgtggacatg ggctgtgccg ggctccggga tgcacacgag gagaaccctg
     2041 agagcatcct ggacgagcac gtacagcgtg tgctgaggac acctggccgc cagtcgcctg
     2101 ggcctggcca tcgctccccg gacagtgggc acgtggccaa gatgccagtg gcactggggg
     2161 gtgccgcctc ggggcacggg aagcacgtac ccaagtcagg ggcgaagctg gacgcggccg
     2221 gcctgcacca ccaccgacac gtccaccacc acgtccacca cagcacagcc cggcccaagg
     2281 agcaggtgga ggccgaggcc acccgcaggg cccagagcag cttcgcctgg ggcctggaac
     2341 cacacagcca tggggcaagg tcccgaggct actcagagag tgttggcgct gcccccaacg
     2401 ccagtgatgg cctcgcccac agtgggaagg tgggcgtggc gtgcaaaaga aatgccaaga
     2461 aggccgagtc ggggaagagc gccagcaccg aggtgccagg tgcctcggag gatgcggaga
     2521 agaaccagaa aatcatgcag tggatcattg agggggaaaa ggagatcagc aggcaccgca
     2581 ggaccggcca cgggtcttcg gggacgagga agccacagcc ccatgagaac tccagaccct
     2641 tgtcccttga gcacccctgg gccggccctc agctccggac ctccgtgcag ccctcccacc
     2701 tcttcatcca agaccccacc atgccacccc acccagctcc caacccccta acccagctgg
     2761 aggaggcgcg ccgacgtctg gaggaggaag aaaagagagc cagccgagca ccctccaagc
     2821 agaggacaag atcgcagagg aaggtgggcg gcgggagtgc ccagccgtgt gacagcatcg
     2881 ttgtggcgta ctacttctgc ggggaaccca tcccctaccg caccctggtg aggggccgcg
     2941 ctgtcaccct gggccagttc aaggagctgc tgaccaaaaa gggcagctac agatactact
     3001 tcaagaaagt gagcgacgag tttgactgtg gggtggtgtt tgaggaggtt cgagaggacg
     3061 aggccgtcct gcccgtcttt gaggagaaga tcatcggcaa agtggagaag gtggactgag
     3121 cggccgctaa gtaagtaagg atccccagct tggccgccat ggcccaactt gtttattgca
     3181 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3241 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     3301 gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     3361 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3421 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3481 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3541 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3601 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3661 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3721 tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3781 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3841 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3901 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3961 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4021 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4081 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4141 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4201 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4261 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     4321 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4381 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4441 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4501 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4561 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4621 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4681 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4801 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4861 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4921 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4981 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5041 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5101 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5161 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5221 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5281 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5341 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5401 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5461 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5521 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5581 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5641 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5701 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5761 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5821 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5881 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5941 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6001 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6061 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6121 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6181 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6241 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6301 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6361 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6421 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6481 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6541 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6601 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6661 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6721 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6781 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
     6841 gattacgaat taattcgagc tcgcccgaca ttgattattg actagttatt aatagtaatc
     6901 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6961 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     7021 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     7081 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     7141 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     7201 tcctacttgg cagtacatct acgtattagt catcgctatt accatg
//
