LOCUS       sequence-7692-ex        5290 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 5290)
  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 5290)
  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-600uDelta4
FEATURES             Location/Qualifiers
     source          1..5290
                     /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(1861..1863,1864..1866,1867..2580)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2636..2654)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2650..2697
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2794..3249
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2881..2900)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3091..3112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3330..3434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3435..3503,3504..4295)
                     /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      4466..5054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4955..4974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5208..5225
                     /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 gaaaggtacc
     1861 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1921 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1981 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2041 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
     2101 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2161 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2221 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2281 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2341 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2401 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2461 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2521 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     2581 aagcttgatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2641 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2701 aaaggaggaa ctatatccgg atctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2761 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2821 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2881 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2941 taaatcgggg gctcccttta gggttccgat ttagagcttt acggcacctc gaccgcaaaa
     3001 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3061 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3121 tcaaccctat cgcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3181 ggttaaaaaa tgagctgatt taacaaatat ttaacgcgaa ttttaacaaa atattaacgt
     3241 ttacaatttc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3301 atacaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3361 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3421 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3481 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3541 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3601 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3661 atacactatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3721 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3781 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3841 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3901 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgaga
     3961 gtgacaccac gatgcctgta gcaatgccaa caacgttgcg caaactatta actggcgaac
     4021 tacttactct agcttcccgg caacaattaa tagactgaat ggaggcggat aaagttgcag
     4081 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4141 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag cgctcccgta
     4201 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4261 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4321 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4381 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4441 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4501 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4561 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4921 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4981 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5041 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctgggc ttttgctggc
     5101 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5161 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5221 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5281 attaatgcag
//
