LOCUS       sequence-8756-ex        7964 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 7964)
  AUTHORS   Takanaga H, Chaudhuri B, Frommer WB
  TITLE     GLUT1 and GLUT9 as major contributors to glucose influx in HepG2 
            cells identified by a high sensitivity intramolecular FRET glucose 
            sensor.
  JOURNAL   Biochim Biophys Acta. 2008 Apr . 1778(4):1091-9.
  PUBMED    18177733
REFERENCE   2  (bases 1 to 7964)
  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: "Biochim 
            Biophys Acta. 2008 Apr . 1778(4):1091-9."
COMMENT     Sequence Label: pEF/myc/ER FLII12Pglu-700uDelta6
FEATURES             Location/Qualifiers
     source          1..7964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..1185
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          254..1176
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1133..1153
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1193..1202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(1199..1243,1326..1337)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(1199..1244,1327..1337)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(1407..1409,1410..1412,1413..2120)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(3018..3020,3021..3023,3024..3734)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3744..3773
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3827..3844)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3833..4057
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4103..4531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4190..4209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4400..4421
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4540..4560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4545..4875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4726..4861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4788..4807
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4896..5690
                     /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(4950..4969)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5560..5579
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5864..5985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5901..5920)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5955..5974
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6016..6033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6187..6775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6267..6286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6946..7737,7738..7806))
                     /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     7569..7588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7807..7911)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgaatt caagcttcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
       61 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      121 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      181 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      241 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      301 tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc ttgatcccga
      361 gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc ttgagttgag
      421 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
      481 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
      541 ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat ttcggttttt
      601 gggcccgcgg ccggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
      661 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
      721 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
      781 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctccagg gggctcaaaa
      841 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
      901 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
      961 ctcgattagt tctggagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     1021 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     1081 atgtaattct ccttggaatt tggccttttt gagtttggat cttggttcat tctcaagcct
     1141 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacg tggccaccat
     1201 gggatggagc tgtatcatcc tcttcttggt agcaacagct acaggtaagg ggttaacagt
     1261 agcaggcttg aggtctggac atatatatgg gtgacaatga catccacttt gcctttctct
     1321 ccacaggcgc gcactcccag gtccaactgc aggtcgacgc catggctgat actcgcattg
     1381 gtgtaacaat ctataagtcg gctggtatgg tgagcaaggg cgaggagctg ttcaccgggg
     1441 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1501 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1561 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc gtgcagtgct
     1621 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1681 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1741 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1801 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc cacaacgtct
     1861 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1921 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1981 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2041 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2101 tcggcatgga cgagctgtac ggtagccgag ataactttat gtctgtagtg cgcaaggcta
     2161 ttgagcaaga tgcgaaagcc gcgccagatg ttcagctgct gatgaatgat tctcagaatg
     2221 accagtccaa gcagaacgat cagatcgacg tattgctggc gaaaggggtg aaggcactgg
     2281 caatcaacct ggttgacccg gcagctgcgg gtacggtgat tgagaaagcg cgtgggcaaa
     2341 acgtgccggt ggttttcttc aacaaagaac cgtctcgtaa ggcgctggat agctacgaca
     2401 aagcctacta cgttggcact gactccaaag agtccggcat tattcaaggc gatttgattg
     2461 ctaaacactg ggcggcgaat cagggttggg atctgaacaa agacggtcag attcagttcg
     2521 tactgctgaa aggtgaaccg ggccatccgg atgcagaagc acgtaccact tacgtgatta
     2581 aagaattgaa cgataaaggc atcaaaactg aacagttaca gttagatacc gcaatgtggg
     2641 acaccgctca ggcgaaagat aagatggacg cctggctgtc tggcccgaac gccaacaaaa
     2701 tcgaagtggt tatcgccaac aacgatgcga tggcaatggg cgcggttgaa gcgctgaaag
     2761 cacacaacaa gtccagcatt ccggtgtttg gcgtcgatgc gctgccagaa gcgctggcgc
     2821 tggtgaaatc cggtgcactg gcgggcaccg tactgaacga tgctaacaac caggcgaaag
     2881 cgacctttga tctggcgaaa aacctggccg atggtaaagg tgcggctgat ggcaccaact
     2941 ggaaaatcga caacaaagtg gtccgcgtac cttatgttgg cgtagataaa gacaacctgg
     3001 ctgaattcag caagaaaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3061 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3121 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3181 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct
     3241 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3301 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3361 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3421 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3481 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3541 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3601 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     3661 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3721 acgagctgta caaggcggcc gcagaacaaa aactcatctc agaagaggat ctgaatgggg
     3781 ccgcaagcga gaaggacgag ctgtagtcta gaagctcgct gatcagcctc gactgtgcct
     3841 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3901 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3961 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4021 aatagcaggc atgctgggga tggcccgggc tctatggctt ctgaggcgga aagaaccagc
     4081 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4141 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4201 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     4261 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4321 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4381 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4441 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     4501 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4561 gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4681 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4741 ccgcccagtt ccgcccattc tccgccccta ggctgactaa ttttttttat ttatgcagag
     4801 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4861 ctaggctttt gcaaaaagct cccgggaggt ccacaatgat tgaacaagat ggattgcacg
     4921 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4981 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5041 tcaagaccga cctgtccggt gccctgaatg aactccagga cgaggcagcg cggctatcgt
     5101 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5161 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5221 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5281 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5341 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5401 aactgttcgc caggctcaag gcgcgtatgc ccgacggcga ggatctcgtc gtgactcatg
     5461 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5521 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5581 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5641 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5701 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5761 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5821 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6001 ggatctttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6061 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6361 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6601 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7921 ccgaaaagtg ccacctgacg tcagatcgac ggatcgggag atcg
//
