LOCUS       sequence-374869-        4763 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression plasmids for Long Stokes shift red fluorescent 
            protein cyScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4763)
  TITLE     Bacterial expression plasmids for Long Stokes shift red fluorescent 
            protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4763)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBAD-cyScarlet
FEATURES             Location/Qualifiers
     source          1..4763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..328
                     /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     251..270
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             374..391
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             395..427
                     /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(431..439,440..454)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             470..1165
                     /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="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGIEADTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(603..621)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     871..890
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1207..1224)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1377..1463
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1555..1582
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1601..1692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1693..1761,1762..2553)
                     /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(1911..1930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2724..3312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3213..3232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3466..3483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3498..3638
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3564..3583)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3724..3746
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(3902..4763,1..17))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca
       61 tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg
      121 gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg
      181 cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc
      241 acactttgct atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt
      301 ttatcgcaac tctctactgt ttctccatac ccgttttttg ggctaacagg aggaattaac
      361 catggggggt tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca
      421 aatgggtcgg gatctgtacg acgatgacga taaggatccg agctcgagaa tggtgagcaa
      481 gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa
      541 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      601 cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc
      661 tcagttcatg tacggctcca gggccttcat caagcacccc gccgacatcc ccgactacta
      721 taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      781 cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa
      841 gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggcat
      901 tgaagcggac accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat
      961 ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc
     1021 caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc
     1081 ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac
     1141 cggcggcatg gacgagctgt acaagtaaaa gcttggctgt tttggcggat gagagaagat
     1201 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
     1261 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
     1321 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     1381 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1441 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     1501 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
     1561 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata
     1621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1921 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2221 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2281 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2401 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2521 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2581 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2761 caaacaaaac aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     2881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3361 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3421 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3481 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3541 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3601 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3661 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3721 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3781 agatcaattc gcgcgcgaag gcgaagcggc atgcataatg tgcctgtcaa atggacgaag
     3841 cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca
     3901 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     3961 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     4021 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     4081 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     4141 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
     4201 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     4261 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
     4321 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     4381 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     4441 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     4501 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     4561 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     4621 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     4681 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     4741 agtatcccgg cagcagggga tca
//
