LOCUS       sequence-7683-ex        5587 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 5587)
  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 5587)
  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
FEATURES             Location/Qualifiers
     source          1..5587
                     /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(202..204,205..207,208..918)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2158..2160,2161..2163,2164..2877)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2933..2951)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2947..2994
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3091..3546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3178..3197)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3388..3409
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3627..3731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3732..3800,3801..4592)
                     /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      4763..5351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5252..5271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5505..5522
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg
      241 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      301 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      361 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
      421 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      481 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      541 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      601 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
      661 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      721 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      781 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      841 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
      901 atggacgagc tgtacaaggg tggtaccgga ggcgccgctg aggcggccga actttccatt
      961 gccgcaaatt cgaccggcaa gaacgtcgcg tttttccgcg aacgcatcgc cgctttcgag
     1021 aaagagaccg gtcacaaggt caatctcgtc accatgccgt cttcttccag cgaacagttc
     1081 agccagtacc ggctgtggct tgccgccggc aacaaggatg tcgacgttta ccagacggac
     1141 gtgatctggg ccccgcagct agccgaacaa ttcgtggatt tgaccgcagc gaccaaggat
     1201 gtcatcgggg atcatttccc ttctatcgtc gcctcacaga cggtggacgg caagctcgtc
     1261 gccatgccga tgttcaccga cgcgccggcg ctgttttacc gcaaggacct tctggaaaaa
     1321 tatggcaagc agccgccgaa aacctggaag gaactgagcg agaccgccaa ggaggttcag
     1381 gacaaggaac gcgcggccgg ccagaaggac ctctggggct tcgtttttca gggcagcgcc
     1441 tatgaaggtc tgacctgcaa cgctctggaa tggatcgcat ccgcgggcgg tggccatatc
     1501 gttgaaacca acggcgatat ctccatcaat aacgagaagg cggcagctgc aatcgaaacc
     1561 gccaaaggct gggtcggcac catcgcaccg cagggcgttc tcgcttacaa ggaagaggaa
     1621 gcacgcggtg tctggcagac cggcaattcc gtcttcatgc gcaactggcc ctatgcctat
     1681 gcgcttggaa atggtgccga cagcgcgatc aaggacaagt tcggcgtcac gccgctgccg
     1741 gcaggtgaag aaggcgcagc ccccgcttcc acactcggcg gcgcgaacct tgctgtttcg
     1801 aaatattccg acgatcagga agcggctatc cagctggtaa aattcctggc atcaaaggat
     1861 acccagaagc tgcgcgcgat ccagctgtcc aacatgccga cgatcgcttc gctttacgac
     1921 gacaaggatg tcgcggccgc acagcccttc atgccgacgt ggaagccgat cttcgagacc
     1981 gccgttccgc gtccttccgc ctcggccaag gtgaaatata acgaggtttc ggccaagttc
     2041 tggggcgccg tgcacaacac gctctccggc aatggtacgg cggcggaaaa tctcgaactt
     2101 ctcgaggttg aactgaccga tctcaaaggc aacggctggg gcgccggtac cggtggaatg
     2161 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2221 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2281 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2341 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     2401 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2461 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2521 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2581 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2641 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2701 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2761 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2821 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaaag
     2881 cttgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     2941 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     3001 ggaggaacta tatccggatc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3061 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3121 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3181 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3241 atcgggggct ccctttaggg ttccgattta gagctttacg gcacctcgac cgcaaaaaac
     3301 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3361 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3421 accctatcgc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3481 taaaaaatga gctgatttaa caaatattta acgcgaattt taacaaaata ttaacgttta
     3541 caatttcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3601 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3661 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3721 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3781 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3841 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3901 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtgata
     3961 cactattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4021 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4081 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4141 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4201 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagagtg
     4261 acaccacgat gcctgtagca atgccaacaa cgttgcgcaa actattaact ggcgaactac
     4321 ttactctagc ttcccggcaa caattaatag actgaatgga ggcggataaa gttgcaggac
     4381 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4441 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagcgc tcccgtatcg
     4501 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4561 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4621 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4681 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4741 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4801 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4861 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4921 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4981 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5041 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5101 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5161 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5221 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5281 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5341 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctgggcttt tgctggcctt
     5401 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5461 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5521 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5581 aatgcag
//
