LOCUS       sequence-374867-        4763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmids for pH sensitive fluorescent protein 
            pHScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4763)
  TITLE     Bacterial expression plasmids for pH sensitive fluorescent proteins
  JOURNAL   Unpublished
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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBAD-pHScarlet
FEATURES             Location/Qualifiers
     source          1..4763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..336
                     /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     259..278
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             382..399
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             403..435
                     /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(439..447,448..462)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             478..1173
                     /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="MVSKGEAVIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGIVADTERLYPEDGVLKGDIK
                     WALRLKDGGRYLADFKTTYKAKKPVQMPGAYNIDRKLVITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(611..629)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     879..898
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1215..1232)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1385..1471
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1563..1590
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1609..1700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1701..1769,1770..2561)
                     /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(1919..1938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2732..3320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3221..3240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3474..3491
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3506..3646
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3572..3591)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3732..3754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(3910..4763,1..25))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca
       61 attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa
      121 ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga
      181 caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc
      241 acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg gatcctacct
      301 gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg ctaacaggag
      361 gaattaacca tggggggttc tcatcatcat catcatcatg gtatggctag catgactggt
      421 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatccgag ctcgagaatg
      481 gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc
      541 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggt
      601 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc ctgggacatc
      661 ctgtcccctc agttcatgta cggctccagg gccttcatca agcaccccgc cgacatcccc
      721 gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      781 gacggcggcg ccgtgaccgt gacccaggac acctccctgg aggacggcac cctgatctac
      841 aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca
      901 atgggcattg tagcggacac cgagcggttg taccccgagg acggcgtgct gaagggcgac
      961 attaagtggg ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc
     1021 tacaaggcca agaagcccgt gcagatgccc ggcgcctaca acatcgaccg taagttggtc
     1081 atcacctccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgagggccgc
     1141 cactccaccg gcggcatgga cgagctgtac aagtaaaagc ttggctgttt tggcggatga
     1201 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     1261 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     1321 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     1381 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1441 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     1501 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     1561 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     1621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1681 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1741 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1801 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1861 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1921 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1981 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2041 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2101 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2161 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2221 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2281 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2341 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2401 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2461 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2521 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2761 gctgcttgca aacaaaacaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3361 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3421 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3481 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3541 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3601 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3661 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3721 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3781 gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat
     3841 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
     3901 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     3961 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     4021 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     4081 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     4141 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     4201 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     4261 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     4321 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     4381 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     4441 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     4501 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     4561 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     4621 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     4681 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     4741 attaaacgag tatcccggca gca
//
