LOCUS       sequence-454425-        4782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 587, Emission:
            610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4782)
  AUTHORS   Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien 
            RY
  TITLE     Improved monomeric red, orange and yellow fluorescent proteins 
            derived from Discosoma sp. red fluorescent protein.
  JOURNAL   Nat Biotechnol. 2004 Dec;22(12):1567-72. Epub 2004 Nov 21.
  PUBMED    15558047
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"; journalName: "Nat 
            Biotechnol."; date: "2004-12"; volume: "22(12)"; pages: "1567-72. 
            Epub 2004 Nov 21"
COMMENT     Sequence Label: mCherry-pBAD
FEATURES             Location/Qualifiers
     source          1..4782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      171..198
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        217..308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(309..377,378..1169)
                     /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(527..546)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1340..1928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1829..1848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2082..2099
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2114..2254
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2180..2199)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2340..2362
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2518..3396)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3423..3707
                     /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     3630..3649
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3753..3770
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3774..3806
                     /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(3810..3818,3819..3833)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             3849..4559
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3994..4012)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4262..4281
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4605..4622)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      join(4775..4782,1..79)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
       61 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
      121 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
      181 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt
      241 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      301 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      361 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      421 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      481 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      541 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      601 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      661 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      721 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      781 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      841 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      901 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      961 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1021 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1081 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1141 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1201 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1261 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1321 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1381 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1441 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1501 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1561 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1621 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1681 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1741 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1801 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1861 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1921 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1981 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2041 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2101 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2161 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     2221 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     2281 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2341 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat
     2401 caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg acgaagcagg
     2461 gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta
     2521 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
     2581 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
     2641 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
     2701 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
     2761 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
     2821 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
     2881 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
     2941 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     3001 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     3061 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     3121 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     3181 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     3241 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     3301 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     3361 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag
     3421 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
     3481 cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
     3541 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
     3601 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
     3661 ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgttttttgg
     3721 gctaacagga ggaattaacc atggggggtt ctcatcatca tcatcatcat ggtatggcta
     3781 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccga
     3841 gctcgagcat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     3901 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     3961 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     4021 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     4081 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     4141 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     4201 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     4261 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     4321 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     4381 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     4441 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     4501 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
     4561 aattcgaagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     4621 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     4681 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     4741 tctccccatg cgagagtagg gaactgccag gcatcaaata aa
//
