LOCUS       sequence-7691-ex        5302 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 5302)
  AUTHORS   Deuschle K, Okumoto S, Fehr M, Looger LL, Kozhukh L, Frommer WB
  TITLE     Construction and optimization of a family of genetically encoded 
            metabolite sensors by semirational protein engineering.
  JOURNAL   Protein Sci. 2005 Sep . 14(9):2304-14.
  PUBMED    16131659
REFERENCE   2  (bases 1 to 5302)
  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: "Protein 
            Sci. 2005 Sep . 14(9):2304-14."
COMMENT     Sequence Label: pRSET FLII32Pglu-2.2m
FEATURES             Location/Qualifiers
     source          1..5302
                     /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 YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1039..1041,1042..1044,1045..1752)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(2648..2666)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2662..2709
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2806..3261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2893..2912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3103..3124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3342..3446
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3447..3515,3516..4307)
                     /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      4478..5066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4967..4986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5220..5237
                     /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 accttcggct acggcctgca gtgcttcgcc
      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 acagccacaa cgtctatatc
      661 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
      721 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      781 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
      841 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
      901 atggacgagc tgtacaaggg tggtaccgga ggcgccgctg atactcgcat tggtgtaaca
      961 atctataagt acgacgataa cgcgatgtct gtagtgcgca aggctattga gcaagatgcg
     1021 aaagccgcgt cggctggtat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1081 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1141 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1201 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc
     1261 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1321 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1381 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1441 ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc
     1501 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     1561 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1621 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     1681 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1741 gacgagctgt acggtagccg agttcagctg ctgatgaatg attctcagaa tgaccagtcc
     1801 aagcagaacg atcagatcga cgtattgctg gcgaaagggg tgaaggcact ggcaatcaac
     1861 ctggttgacc cggcagctgc gggtacggtg attgagaaag cgcgtgggca aaacgtgccg
     1921 gtggttttct tcaacaaaga accgtctcgt aaggcgctgg atagctacga caaagcctac
     1981 tacgttggca ctgactccaa agagtccggc attattcaag gcgatttgat tgctaaacac
     2041 tgggcggcga atcagggttg ggatctgaac aaagacggtc agattcagtt cgtactgctg
     2101 aaaggtgaac cgggccatcc ggatgcagaa gcacgtacca cttacgtgat taaagaattg
     2161 aacgataaag gcatcaaaac tgaacagtta cagttagata ccgcaatgtg ggacaccgct
     2221 caggcgaaag ataagatgga cgcctggctg tctggcccga acgccaacaa aatcgaagtg
     2281 gttatcgcca acaacgatgc gatggcaatg ggcgcggttg aagcgctgaa agcacacaac
     2341 aagtccagca ttccggtgtt tggcgtcgat gcgctgccag aagcgctggc gctggtgaaa
     2401 tccggtgcac tggcgggcac cgtactgaac gatgctaaca accaggcgaa agcgaccttt
     2461 gatctggcga aaaacctggc cgatggtaaa ggtgcggctg atggcaccaa ctggaaaatc
     2521 gacaacaaag tggtccgcgt accttatgtt ggcgtagata aagacaacct ggctgaattc
     2581 agcaagaaat aaaagcttga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     2641 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     2701 ggttttttgc tgaaaggagg aactatatcc ggatctggcg taatagcgaa gaggcccgca
     2761 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     2821 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2881 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2941 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagagct ttacggcacc
     3001 tcgaccgcaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3061 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3121 ctggaacaac actcaaccct atcgcggtct attcttttga tttataaggg attttgccga
     3181 tttcggccta ttggttaaaa aatgagctga tttaacaaat atttaacgcg aattttaaca
     3241 aaatattaac gtttacaatt tcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3301 atttcacacc gcatacaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3361 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3421 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3481 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3541 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3601 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3661 ttctgctatg tgatacacta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3721 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3781 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3841 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3901 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3961 caaacgacga gagtgacacc acgatgcctg tagcaatgcc aacaacgttg cgcaaactat
     4021 taactggcga actacttact ctagcttccc ggcaacaatt aatagactga atggaggcgg
     4081 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4141 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4201 agcgctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4261 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4321 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4381 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4501 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4561 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4621 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4681 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4741 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4801 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4861 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4921 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4981 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5041 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5101 gcttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5161 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5221 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5281 gttggccgat tcattaatgc ag
//
