LOCUS       sequence-158648-        4782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 568, Emission:
            592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4782)
  AUTHORS   Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Hazelwood KL, Davidson 
            MW, Tsien RY
  TITLE     Improving the photostability of bright monomeric orange and red 
            fluorescent proteins.
  JOURNAL   Nat Methods. 2008 Jun;5(6):545-51. doi: 10.1038/nmeth.1209. Epub 
            2008 May 4.
  PUBMED    18454154
REFERENCE   2  (bases 1 to 4782)
  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: 
            "10.1038/nmeth.1209"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "545-51"
COMMENT     Sequence Label: mApple-pBAD
FEATURES             Location/Qualifiers
     source          1..4782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     179..201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(357..1235)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1262..1546
                     /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     1469..1488
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1592..1609
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1613..1645
                     /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(1649..1657,1658..1672)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1688..2398
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2101..2120
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2444..2461)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2614..2700
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2792..2819
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2838..2929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2930..2998,2999..3790)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3148..3167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3961..4549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4450..4469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4703..4720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(4735..4782,1..93)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
       61 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      121 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      181 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc
      241 aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga cgaagcaggg
      301 attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat
      361 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
      421 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
      481 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
      541 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
      601 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
      661 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
      721 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
      781 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
      841 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
      901 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
      961 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     1021 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     1081 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     1141 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     1201 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
     1261 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     1321 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     1381 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     1441 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     1501 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg
     1561 ctaacaggag gaattaacca tggggggttc tcatcatcat catcatcatg gtatggctag
     1621 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatccgag
     1681 ctcgagcatg gtgagcaagg gcgaggagaa taacatggcc atcatcaagg agttcatgcg
     1741 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1801 cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca agggtggccc
     1861 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggtctacat
     1921 taagcaccca gccgacatcc ccgactactt caagctgtcc ttccccgagg gcttcaggtg
     1981 ggagcgcgtg atgaacttcg aggacggcgg cattattcac gttaaccagg actcctccct
     2041 gcaggacggc gtgttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2101 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc gaggagcgga tgtaccccga
     2161 ggacggcgcc ctgaagagcg agatcaagaa gaggctgaag ctgaaggacg gcggccacta
     2221 cgccgccgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2281 catcgtcgac atcaagttgg acatcgtgtc ccacaacgag gactacacca tcgtggaaca
     2341 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaga
     2401 attcgaagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     2461 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     2521 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     2581 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     2641 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     2701 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     2761 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     2821 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2941 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3001 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3061 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3121 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3181 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3241 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3301 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3361 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3421 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3481 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3541 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3601 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3661 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3721 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3781 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3841 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3901 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3961 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4021 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4081 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4141 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4201 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4261 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4321 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4381 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4441 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4501 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4561 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4621 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4681 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     4741 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cg
//
