LOCUS       sequence-202377-        5505 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 5505)
  AUTHORS   Lager I, Looger LL, Hilpert M, Lalonde S, Frommer WB
  TITLE     Conversion of a putative Agrobacterium sugar-binding protein into a 
            FRET sensor with high selectivity for sucrose.
  JOURNAL   J Biol Chem. 2006 Oct 13. 281(41):30875-83.
  PUBMED    16912038
REFERENCE   2  (bases 1 to 5505)
  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: "J Biol 
            Chem. 2006 Oct 13. 281(41):30875-83."
COMMENT     Sequence Label: pRSET FlipSuc-90u delta1
FEATURES             Location/Qualifiers
     source          1..5505
                     /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             69..91
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             111..128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             132..164
                     /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(168..176,177..191)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(204..206,207..209,210..722)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             join(2130..2132,2133..2135,2136..2849)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2905..2923)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2919..2966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3063..3518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3150..3169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3360..3381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3544..3648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3649..3717,3718..4509)
                     /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     complement(3867..3886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4680..5268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5169..5188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5422..5439
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc
      121 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg
      181 atgacgataa ggatccgggc cgcatggtga gcaagggcga ggagctgttc accggggtgg
      241 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      301 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      361 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca
      421 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      481 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      541 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      601 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
      661 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
      721 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
      781 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
      841 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcggtaccg
      901 gaggcgccgc tgaggcggcc gaactttcca ttgccgcaaa ttcgaccggc aagaacgtcg
      961 cgtttttccg cgaacgcatc gccgctttcg agaaagagac cggtcacaag gtcaatctcg
     1021 tcaccatgcc gtcttcttcc agcgaacagt tcagccagta ccggctgtgg cttgccgccg
     1081 gcaacaagga tgtcgacgtt taccagacgg acgtgatctg ggccccgcag ctagccgaac
     1141 aattcgtgga tttgaccgca gcgaccaagg atgtcatcgg ggatcatttc ccttctatcg
     1201 tcgcctcaca gacggtggac ggcaagctcg tcgccatgcc gatgttcacc gacgcgccgg
     1261 cgctgtttta ccgcaaggac cttctggaaa aatatggcaa gcagccgccg aaaacctgga
     1321 aggaactgag cgagaccgcc aaggaggttc aggacaagga acgcgcggcc ggccagaagg
     1381 acctctgggg cttcgttttt cagggcagcg cctatgaagg tctgacctgc aacgctctgg
     1441 aatggatcgc atccgcgggc ggtggccata tcgttgaaac cgacggcgat atctccatca
     1501 ataacgagaa ggcggcagct gcaatcgaaa ccgccaaagg ctgggtcggc accatcgcac
     1561 cgcagggcgt tctcgcttac aaggaagagg aagcacgcgg tgtctggcag accggcaatt
     1621 ccgtcttcat gcgcaactgg ccctatgcct atgcgcttgg aaatggtgcc gacagcgcga
     1681 tcaaggacaa gttcggcgtc acgccgctgc cggcaggtga agaaggcgca gcccccgctt
     1741 ccacactcgg cggcgcgaac cttgctgttt cgaaatattc cgacgatcag gaagcggcta
     1801 tccagctggt aaaattcctg gcatcaaagg atacccagaa gctgcgcgcg atccagctgt
     1861 ccaacatgcc gacgatcgct tcgctttacg acgacaagga tgtcgcggcc gcacagccct
     1921 tcatgccgac gtggaagccg atcttcgaga ccgccgttcc gcgtccttcc gcctcggcca
     1981 aggtgaaata taacgaggtt tcggccaagt tctggggcgc cgtgcacaac acgctctccg
     2041 gcaatggtac ggcggcggaa aatctcgaac ttctcgaggt tgaactgacc gatctcaaag
     2101 gcaacggctg gggcgccggt accggtggaa tggtgagcaa gggcgaggag ctgttcaccg
     2161 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2221 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2281 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt
     2341 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2401 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2461 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2521 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2581 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2641 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2701 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     2761 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2821 ctctcggcat ggacgagctg tacaagtaaa agcttgatcc ggctgctaac aaagcccgaa
     2881 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     2941 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tctggcgtaa
     3001 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3061 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgttggtt cacgtagtgg
     3301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3421 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3481 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     3541 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3601 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3661 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3721 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3781 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3841 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3901 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3961 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4021 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4081 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4141 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4201 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4261 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4321 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4381 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4441 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4501 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4561 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4621 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4681 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4741 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4801 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4861 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4921 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4981 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5041 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5101 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5161 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5221 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5281 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5341 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5401 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     5461 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagg
//
