LOCUS       sequence-477350-        8405 bp    DNA     circular SYN 30-JUN-2025
DEFINITION  Used for transfection of cells for expression of C-MYC EPHA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8405)
  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 8405)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 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 EPHA2
FEATURES             Location/Qualifiers
     source          1..8405
                     /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(3791..3810)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3835..3883
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4085..4513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4172..4191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4382..4403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4522..4542)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4527..4856
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4707..4842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4769..4788
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4923..5717
                     /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(4977..4996)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5587..5606
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5893..6014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5930..5949)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5984..6003
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6063..6079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6063..6079)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6076..6098)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6087..6103
                     /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(6111..6117,6118..6135,6136..6141))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6156..6177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6294..6311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6465..7053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6545..6564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7224..8015,8016..8084))
                     /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     7847..7866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8085..8189)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8264..8283)
                     /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 ggagctccag gcagcccgcg cctgcttcgc cctgctgtgg
      841 ggctgtgcgc tggccgcggc cgcggcggcg cagggcaagg aagtggtact gctggacttt
      901 gctgcagctg gaggggagct cggctggctc acacacccgt atggcaaagg gtgggacctg
      961 atgcagaaca tcatgaatga catgccgatc tacatgtact ccgtgtgcaa cgtgatgtct
     1021 ggcgaccagg acaactggct ccgcaccaac tgggtgtacc gaggagaggc tgagcgtatc
     1081 ttcattgagc tcaagtttac tgtacgtgac tgcaacagct tccctggtgg cgccagctcc
     1141 tgcaaggaga ctttcaacct ctactatgcc gagtcggacc tggactacgg caccaacttc
     1201 cagaagcgcc tgttcaccaa gattgacacc attgcgcccg atgagatcac cgtcagcagc
     1261 gacttcgagg cacgccacgt gaagctgaac gtggaggagc gctccgtggg gccgctcacc
     1321 cgcaaaggct tctacctggc cttccaggat atcggtgcct gtgtggcgct gctctccgtc
     1381 cgtgtctact acaagaagtg ccccgagctg ctgcagggcc tggcccactt ccctgagacc
     1441 atcgccggct ctgatgcacc ttccctggcc actgtggccg gcacctgtgt ggaccatgcc
     1501 gtggtgccac cggggggtga agagccccgt atgcactgtg cagtggatgg cgagtggctg
     1561 gtgcccattg ggcagtgcct gtgccaggca ggctacgaga aggtggagga tgcctgccag
     1621 gcctgctcgc ctggattttt taagtttgag gcatctgaga gcccctgctt ggagtgccct
     1681 gagcacacgc tgccatcccc tgagggtgcc acctcctgcg agtgtgagga aggcttcttc
     1741 cgggcacctc aggacccagc gtcgatgcct tgcacacgac ccccctccgc cccacactac
     1801 ctcacagccg tgggcatggg tgccaaggtg gagctgcgct ggacgccccc tcaggacagc
     1861 gggggccgcg aggacattgt ctacagcgtc acctgcgaac agtgctggcc cgagtctggg
     1921 gaatgcgggc cgtgtgaggc cagtgtgcgc tactcggagc ctcctcacgg actgacccgc
     1981 accagtgtga cagtgagcga cctggagccc cacatgaact acaccttcac cgtggaggcc
     2041 cgcaatggcg tctcaggcct ggtaaccagc cgcagcttcc gtactgccag tgtcagcatc
     2101 aaccagacag agccccccaa ggtgaggctg gagggccgca gcaccacctc gcttagcgtc
     2161 tcctggagca tccccccgcc gcagcagagc cgagtgtgga agtacgaggt cacttaccgc
     2221 aagaagggag actccaacag ctacaatgtg cgccgcaccg agggtttctc cgtgaccctg
     2281 gacgacctgg ccccagacac cacctacctg gtccaggtgc aggcactgac gcaggagggc
     2341 cagggggccg gcagcaaggt gcacgaattc cagacgctgt ccccggaggg atctggcaac
     2401 ttggcggtga ttggcggcgt ggctgtcggt gtggtcctgc ttctggtgct ggcaggagtt
     2461 ggcttcttta tccaccgcag gaggaagaac cagcgtgccc gccagtcccc ggaggacgtt
     2521 tacttctcca agtcagaaca actgaagccc ctgaagacat acgtggaccc ccacacatat
     2581 gaggacccca accaggctgt gttgaagttc actaccgaga tccatccatc ctgtgtcact
     2641 cggcagaagg tgatcggagc aggagagttt ggggaggtgt acaagggcat gctgaagaca
     2701 tcctcgggga agaaggaggt gccggtggcc atcaagacgc tgaaagccgg ctacacagag
     2761 aagcagcgag tggacttcct cggcgaggcc ggcatcatgg gccagttcag ccaccacaac
     2821 atcatccgcc tagagggcgt catctccaaa tacaagccca tgatgatcat cactgagtac
     2881 atggagaatg gggccctgga caagttcctt cgggagaagg atggcgagtt cagcgtgctg
     2941 cagctggtgg gcatgctgcg gggcatcgca gctggcatga agtacctggc caacatgaac
     3001 tatgtgcacc gtgacctggc tgcccgcaac atcctcgtca acagcaacct ggtctgcaag
     3061 gtgtctgact ttggcctgtc ccgcgtgctg gaggacgacc ccgaggccac ctacaccacc
     3121 agtggcggca agatccccat ccgctggacc gccccggagg ccatttccta ccggaagttc
     3181 acctctgcca gcgacgtgtg gagctttggc attgtcatgt gggaggtgat gacctatggc
     3241 gagcggccct actgggagtt gtccaaccac gaggtgatga aagccatcaa tgatggcttc
     3301 cggctcccca cacccatgga ctgcccctcc gccatctacc agctcatgat gcagtgctgg
     3361 cagcaggagc gtgcccgccg ccccaagttc gctgacatcg tcagcatcct ggacaagctc
     3421 attcgtgccc ctgactccct caagaccctg gctgactttg acccccgcgt gtctatccgg
     3481 ctccccagca cgagcggctc ggagggggtg cccttccgca cggtgtccga gtggctggag
     3541 tccatcaaga tgcagcagta tacggagcac ttcatggcgg ccggctacac tgccatcgag
     3601 aaggtggtgc agatgaccaa cgacgacatc aagaggattg gggtgcggct gcccggccac
     3661 cagaagcgca tcgcctacag cctgctggga ctcaaggacc aggtgaacac agtggggatc
     3721 cccatcctcg agccatggaa gcttgatatc taactgactg aaccggtggt accaaacggg
     3781 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3841 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3901 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3961 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4021 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     4081 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4141 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4201 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4261 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4321 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4381 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4441 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4501 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4561 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4621 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4681 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4741 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4801 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4861 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4921 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4981 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5041 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5101 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5161 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5221 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5281 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5341 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5401 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5461 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5521 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5581 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5641 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5701 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     5761 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5821 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5881 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5941 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6001 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6061 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6121 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6181 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6241 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6301 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6361 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6421 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6481 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6541 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6601 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6661 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6721 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6781 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6841 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6901 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6961 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7021 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7081 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7141 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7201 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7261 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7321 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7381 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7441 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7501 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7561 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7621 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7681 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7741 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7801 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7861 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7921 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7981 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8041 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8101 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8161 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8221 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     8281 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     8341 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     8401 ttagg
//
