LOCUS       sequence-477889-        7477 bp    DNA     circular SYN 08-JUL-2025
DEFINITION  Used for transfection of cells for expression of N-MYC KGA-WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7477)
  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 7477)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 8, 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 C-MYC KGA-WT
FEATURES             Location/Qualifiers
     source          1..7477
                     /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"
     CDS             2788..2817
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2863..2882)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2907..2955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3157..3585
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3244..3263)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3454..3475
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3594..3614)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3599..3928
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3779..3914
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3841..3860
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3995..4789
                     /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(4049..4068)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4659..4678
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4965..5086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5002..5021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5056..5075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5135..5151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5135..5151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5148..5170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5159..5175
                     /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(5183..5189,5190..5207,5208..5213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5228..5249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5366..5383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5537..6125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5617..5636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6296..7087,7088..7156))
                     /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     6919..6938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7157..7261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7336..7355)
                     /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 ctctagagaa ttcggatcca tgatgcggct gcgaggctcg
      781 gggatgctgc gggacctgct cctgcggtcg cccgcaggag tatcggcgac tctgcggcgg
      841 gcacagccct tggtcaccct gtgccggcgt ccccgaggcg ggggacggcc ggccgcgggc
      901 ccggctgccg ccgcgcgact ccacccgtgg tggggcgggg gcggctggcc ggcggagccc
      961 ctcgcgcggg gcctgtccag ctctccttcg gagatcttgc aggagctggg caaggggagc
     1021 acgcatccgc agcccggggt gtcgccaccc gctgccccgg cggcgcccgg ccccaaggac
     1081 ggccccgggg agacggacgc gtttggcaac agcgagggca aagagctggt ggcctcaggt
     1141 gaaaataaaa taaaacaggg tctgttacct agcttggaag atttgctgtt ctatacaatt
     1201 gctgaaggac aagagaaaat acctgttcat aaatttatta cagcactcaa atctacagga
     1261 ttgcgaacgt ctgatcccag gttaaaagag tgtatggata tgttaagatt aactcttcaa
     1321 acaacatcag atggtgtcat gctagacaaa gatcttttta aaaaatgtgt tcagagcaac
     1381 attgttttgt tgacacaagc atttagaaga aagtttgtga ttcctgactt tatgtctttt
     1441 acctcacaca ttgatgagtt atatgaaagt gctaaaaagc agtctggagg aaaggttgca
     1501 gattatattc ctcaactggc caaattcagt cccgatttgt ggggtgtgtc tgtttgtaca
     1561 gtagatggac agaggcattc tactggagat accaaagttc ccttctgtct tcagtcctgt
     1621 gtaaaacctt tgaaatatgc cattgctgtt aatgatcttg gaactgaata tgtgcatcga
     1681 tatgttggaa aagagccgag tggactaaga ttcaacaaac tatttttgaa tgaagatgat
     1741 aaaccacata atcctatggt aaatgctgga gcaattgttg tgacttcact aataaagcaa
     1801 ggagtaaata atgctgaaaa atttgactat gtcatgcagt ttttgaataa gatggctggt
     1861 aatgaatatg ttggattcag taatgcaacg tttcagtctg aaagagaaag tggagatcga
     1921 aattttgcaa taggatatta cttaaaagaa aagaagtgtt ttccagaagg cacagacatg
     1981 gttggtatat tagacttcta cttccagctg tgctccattg aagtgacttg tgaatcagcc
     2041 agtgtgatgg ctgcaacact ggctaatggt ggtttctgcc caattactgg tgaaagagta
     2101 ctgagccctg aagcagttcg aaatacattg agtttgatgc attcctgtgg catgtatgac
     2161 ttctcagggc agtttgcttt ccatgttggt cttcctgcaa aatctggagt tgctgggggc
     2221 attcttttag ttgtccccaa tgttatgggt atgatgtgct ggtctcctcc tctggataag
     2281 atgggcaaca gtgttaaggg aattcacttt tgtcacgatc ttgtttctct gtgtaatttc
     2341 cataactatg ataatttgag acactttgca aaaaaacttg atcctcgaag agaaggtggt
     2401 gatcaaaggg taaagtcagt gataaatctt ttgtttgctg catatactgg agatgtgtct
     2461 gcacttcgaa gatttgcttt gtcagctatg gacatggaac agcgggacta tgattctaga
     2521 acagcactcc atgtagctgc tgcagagggt catgttgaag ttgttaaatt tttgctggaa
     2581 gcctgcaaag taaacccttt ccccaaggac aggtggaata acactcccat ggatgaagca
     2641 ctgcactttg gacaccatga tgtatttaaa attctccaag aataccaagt ccagtacaca
     2701 cctcaaggag attctgacaa cgggaaggaa aatcaaacag tccataagaa tcttgatgga
     2761 ttgttgctcg agccatggaa gcttatggaa caaaaactca tctcagaaga ggatctggat
     2821 atctaactga ctgaaccggt ggtccaaacg ggggaggcta actgaaacac ggaaggagac
     2881 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     2941 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     3001 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     3061 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     3121 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3181 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3241 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3301 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3361 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3421 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3481 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3541 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3601 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3661 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3721 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3781 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3841 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     3901 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     3961 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4021 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4081 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4141 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4201 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4261 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4321 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4381 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4441 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4501 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4561 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4621 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4681 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4741 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4801 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4861 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4921 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4981 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5041 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5101 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5161 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5221 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5281 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5941 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7261 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     7321 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     7381 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     7441 aggcttgacc gacaattgca tgaagaatct gcttagg
//
