LOCUS       sequence-158646-        4983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 549, Emission:
            565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4983)
  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 4983)
  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: mOrange2-pBAD
FEATURES             Location/Qualifiers
     source          1..4983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      112..139
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        158..249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(250..318,319..1110)
                     /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(468..487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1281..1869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1770..1789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2023..2040
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2055..2195
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2121..2140)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2281..2303
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2441..2460
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(2640..3518)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3545..3829
                     /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     3752..3771
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             3856..3878
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3898..3915
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3919..3951
                     /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             3964..3984
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3991..4701
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4747..4764)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      join(4917..4983,1..20)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
       61 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
      121 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
      181 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      241 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      301 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      361 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      421 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      481 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      541 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
      601 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
      661 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
      721 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
      781 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
      841 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
      901 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
      961 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1021 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1081 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1141 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1201 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1261 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1321 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1381 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1441 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1501 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1561 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1621 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1681 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1741 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1801 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1861 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1921 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     1981 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2041 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2101 cgcatctggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     2161 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     2221 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2281 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg
     2341 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     2401 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     2461 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     2521 tctgctcatg tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca
     2581 gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat
     2641 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc
     2701 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca
     2761 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg
     2821 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga
     2881 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg
     2941 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt
     3001 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt
     3061 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac
     3121 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat
     3181 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg
     3241 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac
     3301 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga
     3361 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa
     3421 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag
     3481 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc
     3541 agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg
     3601 ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg
     3661 accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac
     3721 attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta
     3781 gcggatacta cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt
     3841 tgggctagaa ataattttgt ttaactttaa gaaggagata tacatatgcg gggttctcat
     3901 catcatcatc atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg
     3961 tacgagaacc tgtacttcca gggctcgagc atggtgagca agggcgagga gaataacatg
     4021 gccatcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac
     4081 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggctttca gaccgctaag
     4141 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcatttc
     4201 acctacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttcaagctg
     4261 tccttccccg agggcttcaa gtgggagcgc gtgatgaact acgaggacgg cggcgtggtg
     4321 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     4381 ggcaccaact tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc
     4441 tcctccgagc ggatgtaccc cgaggacggt gccctgaagg gcaagatcaa gatgaggctg
     4501 aagctgaagg acggcggcca ctacacctcc gaggtcaaga ccacctacaa ggccaagaag
     4561 cccgtgcagc tgcccggcgc ctacatcgtc gacatcaagt tggacatcac ctcccacaac
     4621 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     4681 atggacgagc tgtacaagta agaattcgaa gcttggctgt tttggcggat gagagaagat
     4741 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
     4801 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
     4861 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     4921 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     4981 acg
//
