LOCUS       sequence-177129-        4542 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 4542)
  AUTHORS   Berg J, Hung YP, Yellen G
  TITLE     A genetically encoded fluorescent reporter of ATP:ADP ratio.
  JOURNAL   Nat Methods. 2009 Feb;6(2):161-6. doi: 10.1038/nmeth.1288.
  PUBMED    19122669
REFERENCE   2  (bases 1 to 4542)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2009-02"; volume: "6"; issue: 
            "2"; pages: "161-6"
COMMENT     Sequence Label: pRsetB-his7-Perceval
FEATURES             Location/Qualifiers
     source          1..4542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     200..219
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        200..218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             250..272
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             286..306
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             508..759
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSFQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     696..717
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(820..841)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1081..1100)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2123..2141)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2137..2184
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2281..2736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2368..2387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2578..2599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2762..2866
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2867..2935,2936..3727)
                     /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(3085..3104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3898..4486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4387..4406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      241 agaaataatt ttgtttaact ttaagaagga gatatacata tgaaacatca ccatcaccat
      301 caccatggag ctagcatgaa aaaggtggaa tccatcatca ggcccgaaaa gctggagatc
      361 gttaagaagg ctctctcgga cgctggatat gtgggtatga cagtctctga ggtcaagggc
      421 acgggcgtcc agggcggcat cgtcgagagg taccgaggaa gggagtactc tgcaggctac
      481 aacagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
      541 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
      601 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cttccagtcc
      661 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      721 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggttccgg aggcatggtg
      781 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      841 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      901 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      961 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1021 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1081 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1141 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1201 gagtacaacg gcaccatagt agatctgatc cctaaggtaa aaattgagct cgtggtgaag
     1261 gaggaggacg tcgataacgt gatagacatt atttgcgaaa atgcccgcac aggtaacccc
     1321 ggcgacggta agatatttgt gatcccagtg gagcgagtgg tcagggtgcg aaccaaagag
     1381 gagggagcat ctggtggtgg atccggtgga ggaggtgcgt ctggtatgaa aaaggtggaa
     1441 gctatcatca gacccgaaaa gctggagatc gttaaaaagg ctctttccga cgccggatat
     1501 gtgggtatga cagtctctga ggtcaaaggc cgcggtgcag gtggagggga tctgatccct
     1561 aaggtaaaaa ttgagctcgt ggtgaaggag gaggacgtcg ataacgtgat agacattatt
     1621 tgcgaaaatg cccgcacagg taaccccggc gacggtaaga tatttgtgat cccagtggag
     1681 cgagtggtca gggtgcgaac caaagaggag ggagcatctg gtggaggtgg cggttccgga
     1741 ggtgcatctg gtatgaaaaa ggtggaagct atcatcagac ccgaaaagct ggagatcgtt
     1801 aaaaaggctc tttccgacgc cggatatgtg ggtatgacag tctctgaggt caaaggccgc
     1861 ggtgcaggtg gaggggatct gatccctaag gtaaaaattg agctcgtggt gaaggaggag
     1921 gacgtcgata acgtgataga cattatttgc gaaaatgccc gcacaggtaa ccccggcgac
     1981 ggtaagatat ttgtgatccc agtggagcga gtggtcaggg tgcgaaccaa agaggaggga
     2041 aaggaagcat tgtgactcga ggggcccaag cttgatccgg ctgctaacaa agcccgaaag
     2101 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2161 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatc tggcgtaata
     2221 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     2281 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2341 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2401 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2461 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2521 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2581 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2641 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2701 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     2761 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2821 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2881 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2941 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3001 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3061 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3121 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3181 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3241 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3301 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3361 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3421 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3481 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3541 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3601 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3661 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3721 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3781 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4501 ggccttttta cggttcctgg ccttttgctg gccttttgct ca
//
