LOCUS       sequence-79333-e        6162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of eGFP-fused mouse sedoheptulokinase (CARKL, SHPK).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6162)
  AUTHORS   Kruger A, Gruning NM, Wamelink MM, Kerick M, Kirpy A, Parkhomchuk D,
            Bluemlein K, Schweiger MR, Soldatov A, Lehrach H, Jakobs C, Ralser M
  TITLE     The pentose phosphate pathway is a metabolic redox sensor and 
            regulates transcription during the antioxidant response.
  JOURNAL   Antioxid Redox Signal. 2011 Jul 15;15(2):311-24. doi: 
            10.1089/ars.2010.3797. Epub 2011 May 19.
  PUBMED    21348809
REFERENCE   2  (bases 1 to 6162)
  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"; doi: 
            "10.1089/ars.2010.3797"; journalName: "Antioxid Redox Signal"; date:
            "2011-07-15- 15"; volume: "15"; issue: "2"; pages: "311-24"
COMMENT     Sequence Label: peGFPC2-CARKL
FEATURES             Location/Qualifiers
     source          1..6162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2950..3071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2987..3006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3041..3060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3078..3533)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3215..3236)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3427..3446
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3560..3664
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3666..4023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3666..3857
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3689..3709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3874..4009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4058..4852
                     /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             4058..4852
                     /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(4112..4131)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4722..4741
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5040..5059)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5084..5131
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5460..6048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5949..5968
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agctcaagct tcgaattcat ggcttcgcga
     1381 cctgtcactc tcggtatcga cttgggcacc acgtccgtga aggccgccct attggaggct
     1441 gcacccagcc tcccatccgg gttcgtggtg ctggcgagct gcgcccgggc tgcgagggca
     1501 gagactgaga gcgcggtggc cgggccccag gggcgggagc aggatgtgac tagaatcatc
     1561 caagccctaa atgaatgcct tgatgccctt ccccggcagc agctccagag agtccggggc
     1621 atcggagtgt cgggacagat gcatggcatc ctattttgga aagcaggcca aggctgtgaa
     1681 tggatggagg gagggcctgc ctttgtattt gagcccagag ctgtgagcca cttggtcaca
     1741 tggcaggatg gccgttgtaa cagtagtttc ctggcttctc tgcccaagcc agattcccat
     1801 ctcagtgtgg ccacaggatt cggctgtgca accatcttct ggcttttgaa aaacagcccg
     1861 gagtttctga agtcctatga tgcagctggc accattcaag attatgtggt tgccatgctg
     1921 tgtggcttgc caagacccct gatgtctgac cagaatgccg ctagctgggg ctattttaac
     1981 acccagagcc agagctggaa cttggacaca ctggaaaagg caggctttcc tatccacctt
     2041 ctccccgaca ttgcagagcc aggcagcatg gcaggcagaa cttctcacac atggtttgaa
     2101 atcccaaagg ggacacaggt gggcatagcc ctgggtgact tacaagcctc tgtctattcc
     2161 tgcatgggcc aaaggactga tgcagttctc aacatcagca cctcggtcca gctggcagcc
     2221 tccatgcctg tgggattcca gccactgcag actccggatc ctgcagcccc agttgccttc
     2281 ttcccttact tcgacaggac ctacctaggg gtggcagcgt cactcaatgg aggcaatgtg
     2341 ctggccacgt ttgtccacat gcttgttcag tggatggcag atcttggtct ggaggttgaa
     2401 gaatccactg tgtattcacg catgatccag gcggctgcac agcagaaaga cacccatctc
     2461 accatcaccc caacagtgct gggggagagg cacctgccag accagctggc ctctgtgact
     2521 agaatctcct cctctgacct ctccctgggc catgtgaccc gggccctctg cagaggcatc
     2581 gttcagaacc tgcactccat gcttcctttc caacagctca aagagtgggg tgtggcccgg
     2641 gtggtgggga gtgggagtgc actgtccagg aacgaggtgc tgaagcagga ggtccagagg
     2701 gcgtttcctt tcccagtgtg ctttgggcag gatgtagatg cagcctttgg ggcagctctg
     2761 gtcatgctcc agagagacct tagccagaag gagccttagg tcgacggtac cgcgggcccg
     2821 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2881 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2941 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3001 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3061 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3121 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3181 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3241 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3301 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3361 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3421 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3481 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3541 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3601 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3661 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3721 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3781 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3841 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3901 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3961 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4021 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4081 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4141 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4201 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4261 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4321 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4381 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4441 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4501 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4561 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4621 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4681 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4741 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4801 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4861 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4921 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4981 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5041 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5101 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5161 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5221 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5281 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5341 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5401 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5461 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5521 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5581 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5641 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5701 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5761 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5821 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5881 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5941 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6001 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6061 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6121 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
