LOCUS       sequence-345523-        5151 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 5151)
  AUTHORS   Lager I, Fehr M, Frommer WB, Lalonde S
  TITLE     Development of a fluorescent nanosensor for ribose.
  JOURNAL   FEBS Lett. 2003 Oct 9. 553(1-2):85-9.
  PUBMED    14550551
REFERENCE   2  (bases 1 to 5151)
  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: "FEBS Lett. 
            2003 Oct 9. 553(1-2):85-9."
COMMENT     Sequence Label: FLIPrbs F16A
FEATURES             Location/Qualifiers
     source          1..5151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /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(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(186..188,189..191,192..902)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1758..1760,1761..1763,1764..2477)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2533..2551)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2547..2594
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2691..3146
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2778..2797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2988..3009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3172..3276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3277..3345,3346..4137)
                     /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(3495..3514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4308..4896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4797..4816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5050..5067
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccgg
      181 gccgcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      241 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      301 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      361 ccaccctcgt gaccaccctg acctggggcg tgcagtgctt cagccgctac cccgaccaca
      421 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      481 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      541 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      601 ggcacaagct ggagtacaac tacatcagcc acaacgtcta tatcaccgcc gacaagcaga
      661 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
      721 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      781 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
      841 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
      901 agggtggtac cggaggcgcc atggcaaaag acaccatcgc gctggtggtc tccacgctta
      961 acaacccgtt cgcggtatcg ctgaaagatg gcgcgcagaa agaggcggat aaacttggct
     1021 ataacctggt ggtgctggac tcccagaaca acccggcgaa agagctggcg aacgtgcagg
     1081 acttaaccgt tcgcggcaca aaaattctgc tgattaaccc gaccgactcc gacgcagtgg
     1141 gtaatgctgt gaagatggct aaccaggcga acatcccggt tatcactctt gaccgccagg
     1201 caacgaaagg tgaagtggtg agccacattg cttctgataa cgtactgggc ggcaaaatcg
     1261 ctggtgatta catcgcgaag aaagcgggtg aaggtgccaa agttatcgag ctgcaaggca
     1321 ttgctggtac atccgcagcc cgtgaacgtg gcgaaggctt ccagcaggcc gttgctgctc
     1381 acaagtttaa tgttcttgcc agccagccag cagattttga tcgcattaaa ggtttgaacg
     1441 taatgcagaa cctgttgacc gctcatccgg atgttcaggc tgtattcgcg cagaatgatg
     1501 aaatggcgct gggcgcgctg cgcgcactgc aaactgccgg taaatcgggt gtgatggtcg
     1561 tcggatttga cggtacaccg gatggcgaaa aagcggtgaa tgatggcaaa ctagcagcga
     1621 ctatcgctca gctacccgat cagattggcg cgaaaggcgt cgaaaccgca gataaagtgc
     1681 tgaaaggcga gaaagttcag gctaagtatc cggttgatct gaaactggtt gttaagcagg
     1741 gcgccggtac cggtggaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1801 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1861 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1921 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct
     1981 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2041 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2101 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2161 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     2221 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2281 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2341 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     2401 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2461 acgagctgta caagtaaaag cttgatccgg ctgctaacaa agcccgaaag gaagctgagt
     2521 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     2581 tgaggggttt tttgctgaaa ggaggaacta tatccggatc tggcgtaata gcgaagaggc
     2641 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     2701 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2761 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2821 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2881 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2941 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3001 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3061 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3121 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     3181 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3241 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3301 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3361 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3421 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3481 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3541 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3601 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3661 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3721 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3781 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3841 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3901 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3961 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4021 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4081 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4141 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4201 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4441 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4921 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4981 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5041 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     5101 ctccccgcgc gttggccgat tcattaatgc aggatctcga tcccgcgaaa t
//
