LOCUS       sequence-313841-        7455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7455)
  AUTHORS   Sun L, Shukair S, Naik TJ, Moazed F, Ardehali H
  TITLE     Glucose phosphorylation and mitochondrial binding are required for 
            the protective effects of hexokinases I and II.
  JOURNAL   Mol Cell Biol. 2008 Feb . 28(3):1007-17.
  PUBMED    18039843
REFERENCE   2  (bases 1 to 7455)
  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"; journalName: "Mol Cell 
            Biol. 2008 Feb . 28(3):1007-17."
COMMENT     Sequence Label: FLHKII-pGFPN3
FEATURES             Location/Qualifiers
     source          1..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     RBS             3031..3039
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      3335..3344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3341..3343,3344..3346,3347..4060)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3341..3343,3344..3346,3347..4057)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3386..3407)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3647..3666)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3994..4015
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4183..4304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4220..4239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4274..4293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4311..4766)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4448..4469)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4660..4679
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4793..4897
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4899..5256
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4899..5090
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4922..4942)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5107..5242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5291..6085
                     /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             5291..6085
                     /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(5345..5364)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5955..5974
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6273..6292)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6317..6364
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6693..7281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7182..7201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatgatt gcctcgcatc tgcttgccta
      601 cttcttcacg gagctcaacc atgaccaagt gcagaaggtt gaccagtatc tctaccacat
      661 gcgcctctct gatgagaccc tcttggagat ctctaagcgg ttccgcaagg agatggagaa
      721 agggcttgga gccaccactc accctactgc agcagtgaag atgctgccca cctttgtgag
      781 gtccactcca gatgggacag aacacggaga gttcctggct ctggatcttg gagggaccaa
      841 cttccgtgtg ctttgggtga aggtaacgga caatgggctc cagaaggtgg agatggagaa
      901 tcagatctat gccatccctg aggacatcat gcgaggcagt ggcacccagc tgtttgacca
      961 cattgccgaa tgcctggcta acttcatgga taagctacaa atcaaagaca agaagctccc
     1021 actgggtttt accttctcgt tcccctgcca ccagactaaa ctagacgaga gtttcctggt
     1081 ctcatggacc aagggattca agtccagtgg agtggaaggc agagacgttg tggctctgat
     1141 ccggaaggcc atccagagga gaggggactt tgatatcgac attgtggctg tggtgaatga
     1201 cacagttggg accatgatga cctgtggtta tgatgaccac aactgtgaga ttggtctcat
     1261 tgtgggcacg ggcagcaacg cctgctacat ggaagagatg cgccacatcg acatggtgga
     1321 aggcgatgag gggcggatgt gtatcaatat ggagtggggg gccttcgggg acgatggctc
     1381 gctcaacgac attcgcactg agtttgacca ggagattgac atgggctcac tgaacccggg
     1441 aaagcaactg tttgagaaga tgatcagtgg gatgtacatg ggggagctgg tgaggcttat
     1501 cctggtgaag atggccaagg aggagctgct ctttgggggg aagctcagcc cagagcttct
     1561 caacaccggt cgctttgaga ccaaagacat ctcagacatt gaaggggaga aggatggcat
     1621 ccggaaggcc cgtgaggtcc tgatgcggtt gggcctggac ccgactcagg aggactgcgt
     1681 ggccactcac cggatctgcc agatcgtgtc cacacgctcc gccagcctgt gcgcagccac
     1741 cctggccgcc gtgctgcagc gcatcaagga gaacaaaggc gaggagcggc tgcgctctac
     1801 tattggggtc gacggttccg tctacaagaa acacccccat tttgccaagc gtctacataa
     1861 gaccgtgcgg cggctggtgc ccggctgcga tgtccgcttc ctccgctccg aggatggcag
     1921 tggcaaaggt gcagccatgg tgacagcagt ggcttaccgg ctggccgatc aacaccgtgc
     1981 ccgccagaag acattagagc atctgcagct gagccatgac cagctgctgg aggtcaagag
     2041 gaggatgaag gtagaaatgg agcgaggtct gagcaaggag actcatgcca gtgcccccgt
     2101 caagatgctg cccacctacg tgtgtgctac cccggacggc acagagaaag gggacttctt
     2161 ggccttggac cttggaggaa caaatttccg ggtcctgctg gtccgtgttc ggaatgggaa
     2221 gtggggtgga gtggagatgc acaacaagat ctacgccatc ccgcaggagg tcatgcacgg
     2281 caccggggac gagctctttg accacattgt ccagtgcatc gcggacttcc tcgagtacat
     2341 gggcatgaag ggcgtgtccc tgcctctggg ttttaccttc tccttcccct gccagcagaa
     2401 cagcctggac gagagcatcc tcctcaagtg gacaaaaggc ttcaaggcat ctggctgcga
     2461 gggcgaggac gtggtgaccc tgctgaagga agcgatccac cggcgagagg agtttgacct
     2521 ggatgtggtt gctgtggtga acgacacagt cggaactatg atgacctgtg gctttgaaga
     2581 ccctcactgt gaagttggcc tcattgttgg cacgggcagc aatgcctgct acatggagga
     2641 gatgcgcaat gtggaactgg tggaaggaga agaggggcgg atgtgtgtga acatggaatg
     2701 gggggccttc ggggacaatg gatgcctaga tgacttccgc acagaatttg atgtggctgt
     2761 ggatgagctt tcactcaacc cgggcaagca gaggttcgag aaaatgatca gtggaatgta
     2821 cctgggtgag attgtccgta acattctcat cgatttcacc aagcgtggac tgctcttccg
     2881 aggccgcatc tcagagcggc tcaagacaag gggcatcttt gaaaccaagt tcttgtctca
     2941 gattgagagt gactgcctgg ccctgctgca agtccgagcc atcctgcaac acttagggct
     3001 tgagagcacc tgtgacgaca gcatcattgt taaggaggtg tgcactgtgg tggcccggcg
     3061 ggcagcccag ctctgtggcg caggcatggc cgctgtggtg gacaggatac gagaaaaccg
     3121 tgggctggac gctctcaaag tgacagtggg tgtggatggg accctctaca agctacatcc
     3181 tcactttgcc aaagtcatgc atgagacagt gaaggacctg gctccgaaat gtgatgtgtc
     3241 tttcctgcag tcagaggatg gcagcgggaa gggggcggcg ctcatcactg ctgtggcctg
     3301 ccgcatccgt gaggctggac agcgaggatc catcgccacc atggtgagca agggcgagga
     3361 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3421 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3481 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3541 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3601 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3661 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3721 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3781 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     3841 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3901 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     3961 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4021 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc
     4081 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4141 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4201 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4261 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     4321 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4381 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4441 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4501 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4561 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4621 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4681 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4741 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4801 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4861 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4921 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4981 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5041 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5101 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5161 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5221 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5281 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5341 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5401 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5461 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5521 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5581 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5641 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5701 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5761 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5821 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5881 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5941 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6001 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6061 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6121 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6181 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6241 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6301 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6361 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6421 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6481 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6541 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6601 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6661 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6721 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6781 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6841 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6901 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6961 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7021 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7081 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7141 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7201 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7261 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7321 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7381 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     7441 atatatggag ttccg
//
