LOCUS       sequence-8754-ex        5203 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 5203)
  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 5203)
  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
FEATURES             Location/Qualifiers
     source          1..5203
                     /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(1774..1776,1777..1779,1780..2493)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2549..2567)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2563..2610
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2707..3162
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3004..3025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3243..3347
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3348..3416,3417..4208)
                     /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      4379..4967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4868..4887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5121..5138
                     /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 ggcgccatgg caaaagacac catcgcgctg
      961 gtggtctcca cgcttaacaa cccgttcttt gtatcgctga aagatggcgc gcagaaagag
     1021 gcggataaac ttggctataa cctggtggtg ctggactccc agaacaaccc ggcgaaagag
     1081 ctggcgaacg tgcaggactt aaccgttcgc ggcacaaaaa ttctgctgat taacccgacc
     1141 gactccgacg cagtgggtaa tgctgtgaag atggctaacc aggcgaacat cccggttatc
     1201 actcttgacc gccaggcaac gaaaggtgaa gtggtgagcc acattgcttc tgataacgta
     1261 ctgggcggca aaatcgctgg tgattacatc gcgaagaaag cgggtgaagg tgccaaagtt
     1321 atcgagctgc aaggcattgc tggtacatcc gcagcccgtg aacgtggcga aggcttccag
     1381 caggccgttg ctgctcacaa gtttaatgtt cttgccagcc agccagcaga ttttgatcgc
     1441 attaaaggtt tgaacgtaat gcagaacctg ttgaccgctc atccggatgt tcaggctgta
     1501 ttcgcgcaga atgatgaaat ggcgctgggg gcgctgcgcg cactgcaaac tgccggtaaa
     1561 tcggatgtga tggtcgtcgg atttgacggt acaccggatg gcgaaaaagc ggtgaatgat
     1621 ggcaaactag cagcgactat cgctcagcta cccgatcaga ttggcgcgaa aggcgtcgaa
     1681 accgcagata aagtgctgaa aggcgagaaa gttcaggcta agtatccggt tgatctgaaa
     1741 ctggttgtta agcagggcgc cggtaccggt ggaatggtga gcaagggcga ggagctgttc
     1801 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1861 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1921 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     1981 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2041 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2101 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2161 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2221 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2281 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2341 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     2401 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2461 atcactctcg gcatggacga gctgtacaag taaaagcttg atccggctgc taacaaagcc
     2521 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2581 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctggc
     2641 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2701 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2881 gatttagagc tttacggcac ctcgaccgca aaaaacttga tttgggtgat ggttcacgta
     2941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3001 atagtggact cttgttccaa actggaacaa cactcaaccc tatcgcggtc tattcttttg
     3061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3121 tatttaacgc gaattttaac aaaatattaa cgtttacaat ttcgcctgat gcggtatttt
     3181 ctccttacgc atctgtgcgg tatttcacac cgcatacagg tggcactttt cggggaaatg
     3241 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3301 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3361 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3421 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3481 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3541 caatgatgag cacttttaaa gttctgctat gtgatacact attatcccgt attgacgccg
     3601 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3661 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3721 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3781 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3841 cggagctgaa tgaagccata ccaaacgacg agagtgacac cacgatgcct gtagcaatgc
     3901 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3961 taatagactg aatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4021 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4081 cagcactggg gccagatggt aagcgctccc gtatcgtagt tatctacacg acggggagtc
     4141 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4201 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4261 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4321 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4381 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4441 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4501 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4561 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4621 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4681 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4741 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4801 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4861 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4921 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4981 cggccttttt acggttcctg ggcttttgct ggccttttgc tcacatgttc tttcctgcgt
     5041 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5101 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5161 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cag
//
