LOCUS       sequence-374868-        4775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmids for Long Stokes shift red fluorescent 
            protein LSSmApple.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4775)
  AUTHORS   Ejike JO, Sadoine M, Shen Y, Ishikawa Y, Sunal E, Hansch S, Hamacher
            AB, Frommer WB, Wudick MM, Campbell RE, Kleist TJ
  TITLE     A Monochromatically Excitable Green-Red Dual-Fluorophore Fusion 
            Incorporating a New Large Stokes Shift Fluorescent Protein.
  JOURNAL   Biochemistry. 2024 Jan 2;63(1):171-180. doi: 
            10.1021/acs.biochem.3c00451. Epub 2023 Dec 19.
  PUBMED    38113455
REFERENCE   2  (bases 1 to 4775)
  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/acs.biochem.3c00451"; journalName: "Biochemistry"; date: 
            "2024-01-2- 2"; volume: "63"; issue: "1"; pages: "171-180"
COMMENT     Sequence Label: pBAD-LSSmApple
FEATURES             Location/Qualifiers
     source          1..4775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     203..220
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    235..375
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(301..320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     461..483
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(639..1517)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1544..1828
                     /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     1751..1770
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1874..1891
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1895..1927
                     /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(1931..1939,1940..1954)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1970..2680
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFTWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMIF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGLEACEERMYPEDGALK
                     SEYKEWLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2719..2736)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2889..2975
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3067..3094
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3113..3204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3205..3273,3274..4065)
                     /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(3423..3442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4236..4775,1..49)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4725..4744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
       61 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      121 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      181 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      241 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
      301 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
      361 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      421 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
      481 aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg
      541 aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
      601 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
      661 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
      721 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
      781 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
      841 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
      901 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
      961 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     1021 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     1081 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     1141 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     1201 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     1261 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     1321 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     1381 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     1441 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     1501 ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca
     1561 tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg
     1621 gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg
     1681 cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc
     1741 acactttgct atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt
     1801 ttatcgcaac tctctactgt ttctccatac ccgttttttg ggctaacagg aggaattaac
     1861 catggggggt tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca
     1921 aatgggtcgg gatctgtacg acgatgacga taaggatccg agctcgagaa tggtgagcaa
     1981 gggcgaggag aataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     2041 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2101 ggcctttcag accgctaagc tgaaggtgac caagggtggc cccctgccct tcacctggga
     2161 catcctgtcc cctcagttca tgtacggctc caaggtctac attaagcacc cagccgacat
     2221 ccccgactac ttcaagctgt ccttccccga gggcttcagg tgggagcgcg tgatgatctt
     2281 cgaggacggc ggcattattc acgttaacca ggactcctcc ctgcaggacg gcgtgttcat
     2341 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtta tgcagaagaa
     2401 gaccatgggc cttgaggcct gtgaggagcg gatgtacccc gaggacggcg ccctgaagag
     2461 cgagtataag gagtggctga agctgaagga cggcggccac tacgccgccg aggtcaagac
     2521 cacctacaag gccaagaagc ccgtgcagct gccaggcgcc tacatcgtcg acatcaagtt
     2581 ggacatcgtg tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     2641 ccgccactcc accggcggca tggacgagct gtacaagtaa aagcttggct gttttggcgg
     2701 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     2761 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     2821 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     2881 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     2941 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     3001 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     3061 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta
     3121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3421 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     3481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3721 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4261 atctgctgct tgcaaacaaa acaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4381 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4741 ctttatagtc ctgtcgggtt tcgccacctc tgact
//
