LOCUS       sequence-315807-        7183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-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 7183)
  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 7183)
  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: Axin1_HA in pRK5
FEATURES             Location/Qualifiers
     source          1..7183
                     /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     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3423..3449
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3470..3479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3483..3617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3520..3539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3574..3593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3686..4043
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3686..3877
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3709..3729)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3894..4029
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4063..4080)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4063..4079)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4072..4094)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4292..4747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4379..4398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4589..4610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4764..4783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4883..4905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4943..4961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5029..5133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5134..5202,5203..5994)
                     /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(5352..5371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6165..6753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6654..6673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6907..6924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7041..7062
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7077..7082,7083..7100,7101..7107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7115..7131
                     /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     7120..7142
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7139..7155
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7139..7155
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
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 agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattgaatt cgtcgacgcc accatgaata tccaagagca gggtttcccc ttggacctcg
      961 gagcaagttt caccgaagat gctccccgac ccccagtgcc tggtgaggag ggagaactgg
     1021 tgtccacaga cccgaggccc gccagctaca gtttctgctc cgggaaaggt gttggcatta
     1081 aaggtgagac ttcgacggcc actccgaggc gctcggatct ggacctgggg tatgagcctg
     1141 agggcagtgc ctcccccacc ccaccatact tgaagtgggc tgagtcactg cattccctgc
     1201 tggatgacca agatgggata agcctgttca ggactttcct gaagcaggag ggctgtgccg
     1261 acttgctgga cttctggttt gcctgcactg gcttcaggaa gctggagccc tgtgactcga
     1321 acgaggagaa gaggctgaag ctggcgagag ccatctaccg aaagtacatt cttgataaca
     1381 atggcatcgt gtcccggcag accaagccag ccaccaagag cttcataaag ggctgcatca
     1441 tgaagcagct gatcgatcct gccatgtttg accaggccca gaccgaaatc caggccacta
     1501 tggaggaaaa cacctatccc tccttcctta agtctgatat ttatttggaa tatacgagga
     1561 caggctcgga gagccccaaa gtctgtagtg accagagctc tgggtcaggg acagggaagg
     1621 gcatatctgg atacctgccg accttaaatg aagatgagga atggaagtgt gaccaggaca
     1681 tggacgagga cgatggcaga gacgctgctc cccccggaag actccctcag aagctgctcc
     1741 tggagacagc tgccccgagg gtctcctcca gtagacggta cagcgaaggc agagagttca
     1801 ggtatggatc ctggcgggag ccagtcaacc cctattatgt caatgccggc tatgccctgg
     1861 ccccagccac cagtgccaac gacagcgagc agcagagcct gtccagcgat gcagacaccc
     1921 tgtccctcac ggacagcagc gtggatggga tccccccata caggatccgt aagcagcacc
     1981 gcagggagat gcaggagagc gtgcaggtca atgggcgggt gcccctacct cacattcccc
     2041 gcacgtaccg ggtgccgaag gaggtccgcg tggagcctca gaagttcgcg gaggagctca
     2101 tccaccgcct ggaggctgtg cagcgcacgc gggaggccga ggagaagctg gaggagcggc
     2161 tgaagcgcgt gcgcatggag gaggaaggtg aggacggcga tccatcatca gggcccccag
     2221 ggccgtgtca caagctgcct cccgcccccg cttggcacca cttcccgccc cgctgtgtgg
     2281 acatgggctg tgccgggctc cgggatgcac acgaggagaa ccctgagagc atcctggacg
     2341 agcacgtaca gcgtgtgctg aggacacctg gccgccagtc gcctgggcct ggccatcgct
     2401 ccccggacag tgggcacgtg gccaagatgc cagtggcact ggggggtgcc gcctcggggc
     2461 acgggaagca cgtacccaag tcaggggcga agctggacgc ggccggcctg caccaccacc
     2521 gacacgtcca ccaccacgtc caccacagca cagcccggcc caaggagcag gtggaggccg
     2581 aggccacccg cagggcccag agcagcttcg cctggggcct ggaaccacac agccatgggg
     2641 caaggtcccg aggctactca gagagtgttg gcgctgcccc caacgccagt gatggcctcg
     2701 cccacagtgg gaaggtgggc gtggcgtgca aaagaaatgc caagaaggcc gagtcgggga
     2761 agagcgccag caccgaggtg ccaggtgcct cggaggatgc ggagaagaac cagaaaatca
     2821 tgcagtggat cattgagggg gaaaaggaga tcagcaggca ccgcaggacc ggccacgggt
     2881 cttcggggac gaggaagcca cagccccatg agaactccag acccttgtcc cttgagcacc
     2941 cctgggccgg ccctcagctc cggacctccg tgcagccctc ccacctcttc atccaagacc
     3001 ccaccatgcc accccaccca gctcccaacc ccctaaccca gctggaggag gcgcgccgac
     3061 gtctggagga ggaagaaaag agagccagcc gagcaccctc caagcagagg acaagatcgc
     3121 agaggaaggt gggcggcggg agtgcccagc cgtgtgacag catcgttgtg gcgtactact
     3181 tctgcgggga acccatcccc taccgcaccc tggtgagggg ccgcgctgtc accctgggcc
     3241 agttcaagga gctgctgacc aaaaagggca gctacagata ctacttcaag aaagtgagcg
     3301 acgagtttga ctgtggggtg gtgtttgagg aggttcgaga ggacgaggcc gtcctgcccg
     3361 tctttgagga gaagatcatc ggcaaagtgg agaaggtgga cgcggccgca ggaggtggag
     3421 gttaccccta tgatgtgcct gactacgcat gataaggatc cccagcttgg ccgccatggc
     3481 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3541 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3601 cttatcatgt ctggatcggg aattaattcg gcgcagcacc atggcctgaa ataacctctg
     3661 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3721 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3781 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3841 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3901 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3961 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4021 ttggaggcct aggcttttgc aaaaagctgt taacagcttg gcactggccg tcgttttaca
     4081 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     4141 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4201 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4261 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4321 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4381 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4441 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4501 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4561 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4621 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4681 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4741 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4801 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4861 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4921 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4981 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5041 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5101 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5161 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5221 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5281 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5341 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5401 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5461 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5521 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5581 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5641 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5701 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5761 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5821 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5881 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5941 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6001 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6061 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6121 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6181 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6241 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6301 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6361 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6421 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6481 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6541 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6601 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6661 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6721 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6781 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6841 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6901 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6961 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7021 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7081 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7141 ggaaacagct atgacatgat tacgaattaa ttcgagctcg ccc
//
