LOCUS       sequence-14408-e        3513 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 3513)
  AUTHORS   Tantama M, Hung YP, Yellen G
  TITLE     Imaging Intracellular pH in Live Cells with a Genetically Encoded 
            Red Fluorescent Protein Sensor.
  JOURNAL   J Am Chem Soc. 2011 Jul 6. 133(26):10034-7.
  PUBMED    21631110
REFERENCE   2  (bases 1 to 3513)
  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 Am Chem 
            Soc. 2011 Jul 6. 133(26):10034-7."
COMMENT     Sequence Label: pRsetB-pHRed
FEATURES             Location/Qualifiers
     source          1..3513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             17..37
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             join(47..49,50..52,53..712)
                     /codon_start=1
                     /product="red fluorescent protein that serves as a 
                     ratiometric pH sensor (Tantama et al., 2011)"
                     /label=pHRed
                     /note="contains an A213S mutation relative to the parental 
                     mKeima protein"
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEISIARHSLLG
                     "
     primer_bind     complement(825..843)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      839..886
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      983..1438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1070..1089)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1280..1301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1464..1568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1569..1637,1638..2429)
                     /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(1787..1806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2600..3188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3089..3108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3342..3359
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3444..3463
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3444..3462
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             join(3494..3513,1..3)
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 agatatacat atgaaacatc accatcacca tcaccatgga gctagcatgg tttccgttat
       61 cgcaaagcaa atgacctata aagtttatat gtctggcacc gttaacggcc actacttcga
      121 ggttgaaggt gacggcaagg gcaagcctta cgagggcgag cagaccgtta agctgaccgt
      181 taccaaaggc ggcccactgc ctttcgcctg ggatatcctg tcccctcagc tccaatacgg
      241 ctccatccct ttcaccaaat accccgagga catccctgac tactttaagc agagcttccc
      301 agagggctac acctgggagc gctctatgaa cttcgaggac ggcgcagttt gtaccgtttc
      361 caacgacagc tccatccagg gtaactgctt catctacaac gttaagattt ccggcgagaa
      421 cttcccacct aacggcccag ttatgcagaa gaaaacccaa ggctgggagc ccagcaccga
      481 acgcctgttt gcccgcgacg gtatgctgat tggtaacgat tatatggccc tgaaactgga
      541 aggcggcggt cactacctgt gcgagttcaa gagcacttac aaggctaaaa agcctgttcg
      601 catgcctggc cgccacgaaa tcgaccgcaa gctggatgtt acctctcaca accgcgacta
      661 caccagcgtt gagcagtgtg aaatctctat cgcccgccac tctctgctgg ggacaggcaa
      721 ctccgccgac ggtggtggct ctggtggttc tagatgactc gaggggccca agcttgatcc
      781 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
      841 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
      901 tatatccgga tctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      961 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1021 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1081 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1141 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1201 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1261 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1321 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1381 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1441 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1501 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1561 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1621 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1681 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1741 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1801 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1861 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1921 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1981 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2041 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2101 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2161 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2221 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2281 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2341 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2401 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2461 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2521 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3241 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3301 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3361 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3421 gcaggatctc gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc
     3481 ctctagaaat aattttgttt aactttaaga agg
//
