LOCUS       sequence-478109-        6275 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  Mammalian expression of cytosolic mouse CNDP2 mutant E166L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  TITLE     A secondary BHB metabolism pathway linked to energy balance
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCMV6-CNDP2-E166L
FEATURES             Location/Qualifiers
     source          1..6275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2472..2501
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2520..2543
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2563..2579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2563..2579)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2590..3212
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2789..2808)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3361..3949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3441..3460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4278..4325
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4350..4369
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4557..5351)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4557..5351)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4668..4687)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5278..5297
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     enhancer        5521..5712
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5669..5689
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5714..5818)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5845..6273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5932..5951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6142..6163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gtcagccctc aaagctgtct tccagtatat cgatgaaaac caggaccgct
     1081 acgtcaagaa acttgcagaa tgggtggcca tccagagcgt gtccgcatgg ccggagaaga
     1141 gaggagagat cagaaggatg atggaggtgg cagctgccga tgtccagagg ctggggggct
     1201 ccgtggaact ggtggatatt gggaagcaga agctcccaga tggctcggag ataccacttc
     1261 ctcccattct gctgggcaag ctaggcagcg acccccagaa gaaaaccgtg tgcatttacg
     1321 ggcacctgga cgtgcagcct gcggccctgg aggacgggtg ggacagcgag cccttcacct
     1381 tggtggagcg ggaaggcaag ctgtatggga gaggctccac ggacgataag gggccagtgg
     1441 ccgggtggat gaatgccctg gaagcctatc agaagactgg tcaggagatc ccagtcaacc
     1501 tgcggttctg cctggaaggc atgctggagt ctggctccga aggcctggat gagctcattt
     1561 ttgcccagaa agacaagttc ttcaaagatg tggactatgt gtgcatttca gacaattact
     1621 ggctggggaa gaataagccg tgtatcacgt acggcctcag aggcatctgt tacttcttca
     1681 tcgaggtgga atgcagtgac aaagacctcc attctggggt atatggaggc tcagtgcacg
     1741 aggccatgac ggatctcatt tcactgatgg gctgcctggt agacaagaag gggaagatcc
     1801 tcatcccggg catcaacgat gccgtggcgc ctgtgacgga tgaggagcat gcgctctacg
     1861 accacattga cttcgatatg gaagagttcg ccaaggacgt gggggctgag acccttctgc
     1921 acagctgcaa gaaagacatc ctgatgcaca gatggcgata cccgtccctc tccctgcatg
     1981 ggatcgaagg ggccttctcc ggttcagggg ccaagactgt gatccccaga aaggtggtcg
     2041 gcaagttctc catcaggctc gtgccagaca tgataccgga agttgttagc gagcaagttt
     2101 caagctactt gtcaaaaaaa tttgctgaac tgcagagccc caacaagttc aaggtgtaca
     2161 tgggccacgg cgggaagccc tgggtgtctg acttcaacca tcctcattac caggcgggga
     2221 gaagagccct gaagacagtg tttggtgttg agcctgactt gaccagagaa ggtggcagta
     2281 ttcctgtgac cttgaccttt caagaggcca caggcaaaaa tgtcatgcta ttgccagtgg
     2341 ggtcagcaga tgacggtgcc cactcgcaga atgagaagct caacaggctc aactacatag
     2401 aaggaaccaa aatgctggct gcatatctgt atgaggtgtc tcagctgaag aacacgcgta
     2461 cgcggccgct cgagcagaaa ctcatctcag aagaggatct ggcagcaaat gatatcctgg
     2521 attacaagga tgacgacgat aaggtttaaa cggccggccg cggtcatagc tgtttcctga
     2581 acagatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     2641 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     2701 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     2761 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     2821 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     2881 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     2941 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3001 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3061 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
     3121 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
     3181 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcggc gagcggtatc
     3241 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3301 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3361 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3421 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3481 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3541 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3601 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3661 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3721 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3781 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3841 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3901 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3961 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4021 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4081 atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt
     4141 ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac
     4201 gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga
     4261 ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat
     4321 tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc
     4381 cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     4441 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     4501 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     4561 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     4621 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     4681 gccaacgcta tgtcctgata gcgatccgcc acacccagcc ggccacagtc gatgaatcca
     4741 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     4801 agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc
     4861 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     4921 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     4981 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     5041 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5101 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5161 gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5221 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5281 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5341 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag
     5401 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     5461 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg agttaggggc
     5521 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     5581 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     5641 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     5701 tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5761 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5821 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5881 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5941 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     6001 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6061 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6121 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6181 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6241 atgagctgat ttaacaaaaa tttaacgcga atttt
//
