LOCUS       sequence-191732-        3527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein Rosmarinus for constitutive 
            expression in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3527)
  AUTHORS   Molina RS, Tran TM, Campbell RE, Lambert GG, Salih A, Shaner NC, 
            Hughes TE, Drobizhev M
  TITLE     Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than 
            Enhanced Green Fluorescent Protein under Two-Photon Excitation.
  JOURNAL   J Phys Chem Lett. 2017 Jun 15;8(12):2548-2554. doi: 
            10.1021/acs.jpclett.7b00960. Epub 2017 May 25.
  PUBMED    28530831
REFERENCE   2  (bases 1 to 3527)
  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.jpclett.7b00960"; journalName: "J Phys Chem Lett"; 
            date: "2017-06-15- 15"; volume: "8"; issue: "12"; pages: "2548-2554"
COMMENT     Sequence Label: pRSM32
FEATURES             Location/Qualifiers
     source          1..3527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..128
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        291..395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             396..1190
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             396..1190
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(450..469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1060..1079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      1361..1949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1850..1869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2103..2120
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2205..2224
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2205..2223
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2349..2366
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2370..2402
                     /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(2406..2414,2415..2429)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(3179..3197)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3193..3240
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(3337..3527,1..265)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3424..3443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
       61 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
      121 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
      181 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
      241 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
      301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      361 gataaatgct tcaataatat tgaaaaagga agagtatgat tgaacaagat ggattgcacg
      421 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
      481 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
      541 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
      601 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
      661 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
      721 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
      781 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
      841 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
      901 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
      961 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1021 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1081 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1141 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ctgtcagacc
     1201 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1261 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1321 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1381 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1441 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1501 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1561 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1621 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1681 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1741 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1801 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1861 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1921 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1981 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2041 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2101 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     2161 cgcgttggcc gattcattaa tgcaggatct cgatcccgcg aaattaatac gactcactat
     2221 agggagacca caacggtttc cctctagaat caccgagttt attcttgaca cctgatgcga
     2281 tgaatgatat aataggaaag tactgttttg tttaacttta agaaggggat atacatatgc
     2341 ggggttctca tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg
     2401 gtcgggatct gtacgacgat gacgataagg atccgtcagt gataaagagc gtcatgaaaa
     2461 ttaagctgca tatggacggc atcgtgaacg gtcacaaatt catgatcacc ggtgaggggg
     2521 aaggcaagcc atttgagggg actcatacca ttatcctgaa ggtgaaggaa ggaggaccac
     2581 tgcctttcgc atatgatatc ctcactacag cctttcagta cggcaatcgg gtcttcacaa
     2641 aatatcccaa ggatattcct gactacttca aacagtcatt cccggagggc tacagttggg
     2701 agcggtccat gacctttgag gatcagggcg tgtgtaccgt gacctctgac atcaaactgg
     2761 aaggagacta tttcacctac gatatcaggt ttcacggagt caatttcccc gccggcggtc
     2821 ccgtcatgca gaaaaagact ctgcggtggg aaccaagcac cgagaacatg tacgtgcgag
     2881 acggcgtgct ggtaggcgag gtggagagaa ctctgctgct ggagggcaat aaacatcacc
     2941 gctgtaactt cagaacaact tacaaagcaa agaaagaagt agtcctgcca gaataccact
     3001 ttgttgacca ccggatagag atcttggggc atgacaagga ctataacaat gtggttgtgt
     3061 atgaaaacgc agtggctcgg caacaggcct ctaccctgcc aagcaaggcg aagtaggaat
     3121 tcgaagcttg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     3181 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     3241 ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc accgatcgcc
     3301 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3361 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3421 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3481 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacgg
//
