LOCUS       sequence-7693-ex        5284 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 5284)
  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 5284)
  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: pRSET FLII12Pglu-700uDelta6
FEATURES             Location/Qualifiers
     source          1..5284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             67..89
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..162
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(166..174,175..189)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(244..246,247..249,250..957)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(1855..1857,1858..1860,1861..2574)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2630..2648)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2644..2691
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2788..3243
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2875..2894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3085..3106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3324..3428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3429..3497,3498..4289)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4460..5048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4949..4968
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5202..5219
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gataattttg tttaacttta agaaggagat atacatatgc ggggttctca tcatcatcat
      121 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      181 gacgataagg atccgggccg cgctgatact cgcattggtg taacaatcta taagtcggct
      241 ggtatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      301 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      361 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      421 accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg
      481 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      541 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      601 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      661 cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag
      721 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      781 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      841 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
      901 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacggt
      961 agccgagata acgcgatgtc tgtagtgcgc aaggctattg agcaagatgc gaaagccgcg
     1021 ccagatgttc agctgctgat gaatgattct cagaatgacc agtccaagca gaacgatcag
     1081 atcgacgtat tgctggcgaa aggggtgaag gcactggcaa tcaacctggt tgacccggca
     1141 gctgcgggta cggtgattga gaaagcgcgt gggcaaaacg tgccggtggt tttcttcaac
     1201 aaagaaccgt ctcgtaaggc gctggatagc tacgacaaag cctactacgt tggcactgac
     1261 tccaaagagt ccggcattat tcaaggcgat ttgattgcta aacactgggc ggcgaatcag
     1321 ggttgggatc tgaacaaaga cggtcagatt cagttcgtac tgctgaaagg tgaaccgggc
     1381 catccggatg cagaagcacg taccacttac gtgattaaag aattgaacga taaaggcatc
     1441 aaaactgaac agttacagtt agataccgca atgtgggaca ccgctcaggc gaaagataag
     1501 atggacgcct ggctgtctgg cccgaacgcc aacaaaatcg aagtggttat cgccaacaac
     1561 gatgcgatgg caatgggcgc ggttgaagcg ctgaaagcac acaacaagtc cagcattccg
     1621 gtgtttggcg tcgatgcgct gccagaagcg ctggcgctgg tgaaatccgg tgcactggcg
     1681 ggcaccgtac tgaacgatgc taacaaccag gcgaaagcga cctttgatct ggcgaaaaac
     1741 ctggccgatg gtaaaggtgc ggctgatggc accaactgga aaatcgacaa caaagtggtc
     1801 cgcgtacctt atgttggcgt agataaagac aacctggctg aattcagcaa gaaaatggtg
     1861 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1921 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1981 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2041 accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     2101 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2161 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2221 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2281 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2341 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2401 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2461 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2521 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaagctt
     2581 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2641 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     2701 ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     2761 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     2821 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2881 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2941 gggggctccc tttagggttc cgatttagag ctttacggca cctcgaccgc aaaaaacttg
     3001 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3061 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3121 ctatcgcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3181 aaaatgagct gatttaacaa atatttaacg cgaattttaa caaaatatta acgtttacaa
     3241 tttcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacag
     3301 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3361 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3421 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3481 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3541 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3601 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtgatacac
     3661 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3721 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3781 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3841 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3901 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagagtgaca
     3961 ccacgatgcc tgtagcaatg ccaacaacgt tgcgcaaact attaactggc gaactactta
     4021 ctctagcttc ccggcaacaa ttaatagact gaatggaggc ggataaagtt gcaggaccac
     4081 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4141 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagcgctcc cgtatcgtag
     4201 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4261 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4321 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4381 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4441 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4501 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4561 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4621 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4681 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4741 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4801 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4861 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4921 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4981 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5041 tatggaaaaa cgccagcaac gcggcctttt tacggttcct gggcttttgc tggccttttg
     5101 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5161 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5221 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5281 gcag
//
