LOCUS       sequence-477868-        8651 bp    DNA     circular SYN 08-JUL-2025
DEFINITION  Used for transfection of cells for expression of C-MYC CYTSB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8651)
  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 8651)
  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 N-MYC CYTSB
FEATURES             Location/Qualifiers
     source          1..8651
                     /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"
     primer_bind     complement(4037..4056)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4081..4129
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4331..4759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4418..4437)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4628..4649
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4768..4788)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4773..5102
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4953..5088
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5015..5034
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5169..5963
                     /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(5223..5242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5833..5852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6139..6260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6176..6195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6230..6249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6309..6325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6309..6325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6322..6344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6333..6349
                     /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(6357..6363,6364..6381,6382..6387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6402..6423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6540..6557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6711..7299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6791..6810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7470..8261,8262..8330))
                     /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     8093..8112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8331..8435)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8510..8529)
                     /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 gcggagtgca gccaagccct ggagtccagt cgccaaagca
      841 ggcagccatg gtcctgatcg cactcggccc ctccctggca ctccttctgg catgaagagt
      901 tccaagtctt caacctcact ggctttcgag tctcggctga gtaagctcaa gagggccagc
      961 agcgaggaca cactcaacaa acctggaagt gcctcttcag gggtggcgag actgaagaag
     1021 acctcaacct ccggagccat ctctgagctc actgagagcc gcctgaggag caacacaggg
     1081 actattccaa caactaaacg gacaggcatt ccagctccac gggagctctc agtaactatc
     1141 tccagagaga ggtctgtgcc acgtggtccc tccagctcaa agaagttagg ctccagccca
     1201 acttcttcct gcaatcccac tcctaccaag catctgagga ccacccctgc aaaacctaag
     1261 caagagcatg aaggtgcaga gaaggctgtg ctcgagtccc aggtacgaga acttctggca
     1321 gaagccaaaa ccaaagacag tgaaatcaac aggcttcgaa gtgaactaaa aaaatgcaaa
     1381 gagaggtggg ccctgagcac tgaggatgcc aatgcttcag acccaagtgc agagggaaca
     1441 gcatccccag aaagtgatgc acagcctttg ataagaaccc tggaggagaa gaacaagacc
     1501 tttcagaaag agcttgctga cctagaggaa gaaaacaggg ccttgaagga gaaactgact
     1561 taccttgagc agtcaccaaa ttcagaaggg gcagcaagtc acactggtga cagcagctgc
     1621 ccaacatcta taacccacga gtccagtttt ggaagcccag ttggaaatga gttgtctagt
     1681 gaaacagatg agtacagaag aaccacacat gggagtgcct tgcggacatc agggtcttca
     1741 agcagtgatg tcaccaaagc ctctctgtca ccggacgcct cagattttga gcatatcact
     1801 gcagacaccc cgtctcggcc cctgtcagcc actagcaacc cttttaagag ctccaagggc
     1861 tcccctactg ggagctcccc aaacaatgca agtgagctgt ccttggcatc tctcacggag
     1921 aagatacaga agatggagga gaatcagcac agcactgcag aagagctgca ggctactctg
     1981 caggaattat cagaccagca gcagatggtg caggaactga cagccgagaa cgagaagcta
     2041 gtggatgaga agaccattct ggagacctcc ttccaccagc accgagagag ggcagaacag
     2101 ctgagtcagg aaaatgagaa gctgatcaac cttttacagg agcgagtgaa gaatgaggaa
     2161 cccagtgccc agggagggaa ggtccttgag ctggagcaga agtgcacgga tattctcgag
     2221 aagagccgct ttgaaagaga gaagctgctc aacattcagc agcagctgac ctgtagccta
     2281 cggaaggttg aggaagaaaa ccaaggcgct atagacatga ttaagcacct gaaggaagaa
     2341 aatgagaagc tgaatgggtt cctagaacat gagcggtgta ataatagtgt gatggccaaa
     2401 actctggagg aatgtcgagt taccttggaa ggcttgaaaa tggagaatgg gtccttgaag
     2461 gctcttttgg aggctgataa gcaaaaagcc atagaggcca gcagtactgt ggggcagaca
     2521 gcagagaact tcgaggttca ggagatgctg aaagtggctc gagctgagaa agatcagttg
     2581 cagctgtctt gcacagagct cagacaagaa ttgctaaagg caaatggtga aattaaacat
     2641 gtttccagcc tgctagctaa gatggagaaa gattattcgt acctgaaaga agtctgtgat
     2701 catcaagcag aacagctgag cagaaccagc ctaaaattac aagagaaagc ttcagaaagt
     2761 gacgcagaaa tcaaagacat gaaagagacc atatttgaat tggaagacca ggtggagcag
     2821 catcgcgccg tcaagttaca caataaccag ctcatcagtg agctggaagg cagtgtgatc
     2881 aagctggagg agcagaagtc agacctagag cggcagttaa agacgctaac aaaacagata
     2941 aaggaggaaa cagaagaatg gaggcgattc caggcagact tacagactgc agtggtagtg
     3001 gccaacgaca tcaagtgtga ggctcagcag gagctgcgca ctgtgaagag gaggttgctg
     3061 gaggaggaag agaaaaatgc caggctgcag aaggagctgg gagacataca ggggcacagc
     3121 agacctgtga atgaagagcc agagccctca gaggccgatg ctgctggccg gtggcgtgga
     3181 gtctatgtca acagaacatc tccagctcct tcagactctg caaccacagt aaagtcactg
     3241 atcaaatcat ttgacttagg acactcaggt ggaactggcc agagtatttc tgtccacaag
     3301 actcccagaa gccccctgag tggaatccca gtgaggactg ctcctgccgc tgctgtctct
     3361 ccgatgcaga gacattcaac atacagcagc atgaagccag ccagcaaagg gacgtctcag
     3421 cgcttggatc tgccggacct tcccctctct gatctgctga agggaagggc cgaggaccgg
     3481 aagtcagacc cttacctccg aaagagtccc tcactggagt ccctgagcag acccccctcc
     3541 ctgggctttg ggaacaccag actgctgagt gcgtccaccg ggggtctgaa gccaagcaag
     3601 ctcagtgttg aaagaagaga ccctctggca gccctggccc gggagtatgg tggctctaag
     3661 cgtaacgcac tcctgaaatg gtgtcagaag aagacagaag gttatgcgaa cattgatatc
     3721 accaatttca gcagcagctg gagtgatggg ctggccctct gtgctctgct gcacacctac
     3781 ctgcctgccc acatcccata ccaggaattg aacagtcagg agaaaaaaag gaatctcctg
     3841 ttggcatttg aagccgcgca gagtgtaggc atcaacccca gcctggagct cagtgagatg
     3901 ctgtacacgg accggcctga ctggcagagc gtgatgcagt atgtggccca gatctacaag
     3961 tactttgaga cgctcgagcc atggaagctt gatatctaac tgactgaacc ggtggtacca
     4021 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4081 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     4141 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4201 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4261 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     4321 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4381 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4441 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4501 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4561 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4621 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4681 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4741 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4801 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4861 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4921 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4981 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5041 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5101 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     5161 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5221 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5281 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5341 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5401 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5461 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5521 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5581 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5641 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5701 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5761 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5821 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5881 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5941 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     6001 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6061 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6121 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6181 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6241 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6301 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6361 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6421 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6481 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6541 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6601 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6661 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6721 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6781 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6841 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6901 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6961 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7021 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7081 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7141 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7201 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7261 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7381 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7441 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7501 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7561 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7621 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7681 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7741 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7801 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7861 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7921 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7981 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8041 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8101 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8161 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8221 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8281 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8341 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8401 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8461 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     8521 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     8581 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     8641 atctgcttag g
//
