LOCUS       sequence-372015-        4763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mWatermelon GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4763)
  AUTHORS   Gohil K, Wu SY, Takahashi-Yamashiro K, Shen Y, Campbell RE
  TITLE     Biosensor Optimization Using a Forster Resonance Energy Transfer 
            Pair Based on mScarlet Red Fluorescent Protein and an 
            mScarlet-Derived Green Fluorescent Protein.
  JOURNAL   ACS Sens. 2023 Feb 24;8(2):587-597. doi: 10.1021/acssensors.2c01730.
            Epub 2023 Jan 24.
  PUBMED    36693235
REFERENCE   2  (bases 1 to 4763)
  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: 
            "10.1021/acssensors.2c01730"; journalName: "ACS Sens"; date: 
            "2023-02-24- 24"; volume: "8"; issue: "2"; pages: "587-597"
COMMENT     Sequence Label: pBAD-mWatermelon
FEATURES             Location/Qualifiers
     source          1..4763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(121..999)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1026..1310
                     /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     1233..1252
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1356..1373
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1377..1409
                     /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(1413..1421,1422..1436)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1452..2147
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEALIKEYMRFKVHMEGSMDGHEFEIEGEGEGRPYEGTHTA
                     KLKVTKGGPLPFSWDILSPQFGYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIHKVKLRGTNFPPDGPVMQRKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADCKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1585..1603)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2189..2206)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2359..2445
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2537..2564
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2583..2674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2675..2743,2744..3535)
                     /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     complement(2893..2912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3706..4294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4195..4214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4448..4465
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4480..4620
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4546..4565)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4706..4728
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc
       61 agggattctg caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa
      121 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
      181 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
      241 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
      301 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
      361 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
      421 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
      481 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
      541 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
      601 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
      661 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
      721 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
      781 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
      841 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
      901 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
      961 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
     1021 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
     1081 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     1141 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     1201 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
     1261 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
     1321 tgggctaaca ggaggaatta accatggggg gttctcatca tcatcatcat catggtatgg
     1381 ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac gataaggatc
     1441 cgagctcgag aatggtaagc aagggcgagg cattgatcaa ggagtacatg cggttcaagg
     1501 tgcacatgga gggctccatg gacggccacg agttcgagat cgagggcgag ggcgagggcc
     1561 gcccctacga gggcacccat accgccaagc tgaaggtgac caagggtggc cccctgccct
     1621 tctcctggga catcctgtcc cctcagttcg ggtacggctc cagggccttc atcaagcacc
     1681 ccgccgacat ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg
     1741 tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg
     1801 gcaccctgat ccacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa
     1861 tgcagaggaa gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg
     1921 tgctgaaggg cgacattaag atggccctgc gcctgaagga cggcggccgc tacctggcgg
     1981 actgcaagac cacctacaag gccaagaagc ccgtgcagat gcccggcgcc tacaacgtcg
     2041 accgcaagtt ggacatcacc tcccacaacg aggactacac cgtggtggaa cagtacgaac
     2101 gctcagaggg ccgccactcc accggcggca tggacgagct gtacaagtaa aagcttggct
     2161 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     2221 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     2281 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     2341 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2401 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     2461 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     2521 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     2581 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2641 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2701 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2761 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2821 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2881 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     2941 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3001 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3061 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3121 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3181 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3241 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3301 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3361 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3421 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3481 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3541 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3601 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3661 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3721 tctgcgcgta atctgctgct tgcaaacaaa acaaccaccg ctaccagcgg tggtttgttt
     3781 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3841 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3901 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3961 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4021 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4081 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4141 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4201 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4261 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4321 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4381 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4441 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4501 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     4561 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     4621 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4681 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4741 tcaccgaaac gcgcgaggca gca
//
