LOCUS       sequence-477317-        7736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for transfection of cells for expression of C-MYC LIMA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7736)
  AUTHORS   Li M, Wang Y, Wei X, Cai WF, Wu J, Zhu M, Wang Y, Liu YH, Xiong J, 
            Qu Q, Chen Y, Tian X, Yao L, Xie R, Li X, Chen S, Huang X, Zhang C, 
            Xie C, Wu Y, Xu Z, Zhang B, Jiang B, Wang ZC, Li Q, Li G, Lin SY, Yu
            L, Piao HL, Deng X, Han J, Zhang CS, Lin SC
  TITLE     AMPK targets PDZD8 to trigger carbon source shift from glucose to 
            glutamine.
  JOURNAL   Cell Res. 2024 Jun 19. doi: 10.1038/s41422-024-00985-6.
  PUBMED    38898113
REFERENCE   2  (bases 1 to 7736)
  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 Res. 
            2024 Jun 19. doi: 10.1038/s41422-024-00985-6."
COMMENT     Sequence Label: pcDNA 3.3 N-MYC LIMA1
FEATURES             Location/Qualifiers
     source          1..7736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      742..751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             751..780
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1821..1832)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3122..3141)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3166..3214
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3416..3844
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3503..3522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3713..3734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3853..3873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3858..4187
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4038..4173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4100..4119
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4254..5048
                     /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(4308..4327)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4918..4937
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5224..5345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5261..5280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5315..5334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5394..5410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5394..5410)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5407..5429)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5418..5434
                     /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(5442..5448,5449..5466,5467..5472))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5487..5508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5625..5642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5796..6384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5876..5895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6555..7346,7347..7415))
                     /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     7178..7197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7416..7520)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7595..7614)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actgagctcc ttaaggttaa cgccaccatg gaacaaaaac tcatctcaga agaggatctg
      781 tctagagaat tcggatccat ggaatcgact ccatttaata gacgccagtg gacttccctg
      841 tcattgagag taacagccaa agagctttct cttgtcaaca agaacaagtc atccgcaatt
      901 gtggaaatat tctccaagta ccagaaggct gctgaagaag ccaacatgga aaggaagaaa
      961 aataaccctg aaagtctgcc ccagcacttt agaagaggga ccctgtctgt gttaaagaag
     1021 aagtgggaga acccggtggc tggggcagaa ttccacacag actcactgcc aaacagcagc
     1081 agtgagggtg ggcacacagc ggactaccct cctgctgaag tgacggacaa gcctgctcct
     1141 ggagtcagag ctgaccggga agagcacacc cagcccaaac ctagatttgg atctcgtccg
     1201 gaagcagtta tccagagccg gtatcctcgc tcagagaaca gccacgattt taaagcccag
     1261 gccacggaga gccaaaaaat ggaaaactgt ctgggagatt ccaggcatga agcagagaag
     1321 ccagagacga gcgaaaacac agaaacttca ggcaaaatag agaaatacaa cgttccactg
     1381 aatagactga agatgatgtt tgagaaaggt gaacacaacc aaaccaagag tctctggacc
     1441 caaagccgaa atgcgggtgg aaggaggctc tctgaaaaca actgttccct ggatgactgg
     1501 gaaataggtg cgggtcattt gtcttcctct gcattcaact cggagaaaaa cgagagtaag
     1561 aggaatctgg agctgccacg cctctcagaa acctccataa aggaccgcat ggccaagtac
     1621 caggctgcag tgtccaagca gagcagccca gccagctaca caaatgagct gaaaaccagt
     1681 gaaagcaaaa ctcataaatg ggaacagaag gagaatgtgc ccccaggtcc cgaggcctgc
     1741 agcgtccatc aggaaggaag caaggtttct acaactgaga atagtctagt ggccctctct
     1801 gtccctgctg aagatgacac ctgtaactcc caggtgaaga gcgaggccca gcagcctatg
     1861 caccctaagc cgctgagtcc tgacgccagg acctccagtc ttccggaaag ttctccttcc
     1921 aaaacagcga agaagtttca ggcgccggca aaggagagct gcgtggagtg tcagaagacg
     1981 gtgtacccca tggaacggct cctggccaac cagcaggtgt ttcacatcag ctgtttccga
     2041 tgctcctact gcaacaacaa gctcagtcta ggaacatatg catccttaca tggacgaatc
     2101 tactgtaagc ctcacttcaa tcaactcttc aagtccaaag gcaactatga cgagggcttt
     2161 gggcacaagc agcataagga tctgtgggca agcaagagcg acaatgagga gactttgggg
     2221 agaccagccc agccgcctaa tgcaggggag agcccccata gcccgggtgt agaagatgcc
     2281 cccatcgcca aggtcggcgt gctggcggca agtatggaag ccaaggcctc ttctcagagg
     2341 gagagggagg ataagcctgc ggagaccaag aagctgagga tcgcctggcc tccgccagcc
     2401 gagctgggcg gttccggaag tgccctggag gaagggatca aagtatcgaa gcccaagtgg
     2461 cctccggagg atgacgtctg caagacggag gccccggagg atgtagatct ggacctgaag
     2521 aagctgcggc ggtcttcgtc cctgaaggag aggagccgcc cgtttacggt agcagcttcc
     2581 tttcgaacct cttccatcaa gagccccaag gcctcttcgc catctctcag gaaaggttgg
     2641 agcgagtccg agcagagtga ggagtttgga ggagggatag cgacgatgga aaggaaacaa
     2701 acggaaaatg ccagaccctc tggggagaaa gagaatgtgg gaaaatcacg ctggcagggc
     2761 gaggaagtgc cgaggagcaa ggaccgtagt tcttttgagc tggagagtga gaattttatg
     2821 gaaaatggag caaacatagc tgaagatgac aaccatgtcc acgcacagca gtccccgtta
     2881 gaacccgagg ctccgggttg gtctggcttt gtagacacca cggctgctaa agaattcact
     2941 acccagaatc aaaaatccca agatgtcggg ttctgggagg gagaagtggt ccgagagctg
     3001 tctgtggagg aacagataaa gagaaaccgg tactatgatg aggacgagga tgaagaactc
     3061 gagccatgga agcttgatat ctaactgact gaaccggtgg taccaaacgg gggaggctaa
     3121 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     3181 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     3241 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     3301 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     3361 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     3421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3541 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3781 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3841 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3961 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4021 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4081 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4141 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4201 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4261 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4321 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4381 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4441 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4501 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4561 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4621 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4681 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4741 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4801 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4861 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4921 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4981 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5041 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     5101 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5161 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5221 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5281 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5341 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5401 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5461 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5521 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5581 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5641 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5701 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5761 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5821 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5881 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5941 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6001 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6061 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6121 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6181 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6241 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6301 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6361 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6421 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6481 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6541 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6601 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6661 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6721 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6781 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6841 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6901 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6961 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7021 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7081 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7141 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7201 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7261 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7321 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7381 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7441 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7501 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7561 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7621 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     7681 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
