LOCUS       sequence-424652-        4782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of pH-sensitive monomeric mNectarine red 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4782)
  AUTHORS   Johnson DE, Ai HW, Wong P, Young JD, Campbell RE, Casey J
  TITLE     Red fluorescent protein pH biosensor to detect concentrative 
            nucleoside transport.
  JOURNAL   J Biol Chem. 2009 Jul 31;284(31):20499-511. doi: 
            10.1074/jbc.M109.019042.
  PUBMED    19494110
REFERENCE   2  (bases 1 to 4782)
  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: "10.1074/jbc.M109"; 
            journalName: "J Biol Chem"; date: "2009-07-31- 31"; volume: "284"; 
            issue: "31"; pages: "20499-511"
COMMENT     Sequence Label: pBAD-mNectarine
FEATURES             Location/Qualifiers
     source          1..4782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(117..908,909..977))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     740..759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(978..1069)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1088..1115)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1207..1293)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1446..1463
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(1509..2219)
                     /codon_start=1
                     /product="pH-sensitive monomeric red fluorescent protein 
                     derived from DsRed and mTangerine (Johnson et al., 2009)"
                     /label=mNectarine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFCYGSKAYVKHPADIPDYLKLSFPEGLNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTVGWEASTERMHPEDGALK
                     GEIMQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVELYERA
                     EGRHSTGGMDELYK"
     primer_bind     2056..2074
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(join(2235..2249,2250..2258))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(2262..2294)
                     /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             complement(2298..2315)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2361..2645)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(2419..2438)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2672..3550
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(3706..3728)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3814..3954
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3869..3888
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3969..3986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4140..4728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4220..4239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
       61 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
      121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
      181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
      241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
      301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
      361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
      421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
      481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
      541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
      601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
      661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
      721 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
      781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
      841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
      901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
      961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaaa gagtttgtag
     1081 aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc tggcagttta
     1141 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc
     1201 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     1261 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
     1321 ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt
     1381 caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga ccgcttctgc
     1441 gttctgattt aatctgtatc aggctgaaaa tcttctctca tccgccaaaa cagccaagct
     1501 tcgaattctt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcagcgcgt
     1561 tcgtacagtt ccacgatggt gtagtcctcg ttgtgggaaa ggatgtccaa cttgatgtcg
     1621 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     1681 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgca tgatctcgcc cttcagggcg
     1741 ccgtcctcgg ggtgcatccg ctcagtggag gcctcccagc ccacggtacg acactgcatt
     1801 acggggccgt cggaggggaa gttagtgccg cgcagcttca ccttgtagat gaactcgccg
     1861 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     1921 cgctcccaat tgaggccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     1981 tgcttcacat aggccttgga gccgtagcag aactgagggg acaggatgtc ccaggcgaag
     2041 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     2101 ccctcgccct caccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     2161 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg
     2221 ctcgagctcg gatccttatc gtcatcgtcg tacagatccc gacccatttg ctgtccacca
     2281 gtcatgctag ccataccatg atgatgatga tgatgagaac cccccatggt taattcctcc
     2341 tgttagccca aaaaacgggt atggagaaac agtagagagt tgcgataaaa agcgtcaggt
     2401 aggatccgct aatcttatgg ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa
     2461 ataatcaatg tggacttttc tgccgtgatt atagacactt ttgttacgcg tttttgtcat
     2521 ggctttggtc ccgctttgtt acagaatgct tttaataagc ggggttaccg gtttggttag
     2581 cgagaagagc cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg gacaattggt
     2641 ttcttctctg aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct
     2701 gccgggatac tcgtttaatg cccatctggt ggcgggttta acgccgattg aggccaacgg
     2761 ttatctcgat ttttttatcg accgaccgct gggaatgaaa ggttatattc tcaatctcac
     2821 cattcgcggt cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga
     2881 tattttgctg ttcccgccag gagagattca tcactacggt cgtcatccgg aggctcgcga
     2941 atggtatcac cagtgggttt actttcgtcc gcgcgcctac tggcatgaat ggcttaactg
     3001 gccgtcaata tttgccaata cggggttctt tcgcccggat gaagcgcacc agccgcattt
     3061 cagcgacctg tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct attcggagct
     3121 gctggcgata aatctgcttg agcaattgtt actgcggcgc atggaagcga ttaacgagtc
     3181 gctccatcca ccgatggata atcgggtacg cgaggcttgt cagtacatca gcgatcacct
     3241 ggcagacagc aattttgata tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg
     3301 tctgtcacat cttttccgcc agcagttagg gattagcgtc ttaagctggc gcgaggacca
     3361 acgtatcagc caggcgaagc tgcttttgag caccacccgg atgcctatcg ccaccgtcgg
     3421 tcgcaatgtt ggttttgacg atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg
     3481 ggccagcccg agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa
     3541 gttgtcataa ttggtaacga atcagacaat tgacggcttg acggagtagc atagggtttg
     3601 cagaatccct gcttcgtcca tttgacaggc acattatgca tgccgcttcg ccttcgcgcg
     3661 cgaattgatc tgctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3721 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3781 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     3841 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3901 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     3961 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4021 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4081 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4141 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4201 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4261 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4321 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4381 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4441 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4501 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4561 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4621 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4681 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4741 atcttttcta cggggtctga cgctcagtgg aacgaaaact ca
//
