LOCUS       sequence-190519-        4104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KillerOrange gene for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4104)
  AUTHORS   Sarkisyan KS, Zlobovskaya OA, Gorbachev DA, Bozhanova NG, Sharonov 
            GV, Staroverov DB, Egorov ES, Ryabova AV, Solntsev KM, Mishin AS, 
            Lukyanov KA
  TITLE     KillerOrange, a Genetically Encoded Photosensitizer Activated by 
            Blue and Green Light.
  JOURNAL   PLoS One. 2015 Dec 17;10(12):e0145287. doi: 
            10.1371/journal.pone.0145287. eCollection 2015.
  PUBMED    26679300
REFERENCE   2  (bases 1 to 4104)
  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: "PLoS One.";
            date: "2015-12-17"; volume: "10(12):e0145287. doi"; pages: " 
            10.1371/journal.pone.0145287. eCollection 2015"
COMMENT     Sequence Label: KillerOrange /pQE-30
FEATURES             Location/Qualifiers
     source          1..4104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(68..859,860..928))
                     /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     691..710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(929..1033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1101..1119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    1163..1179
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1198..1209
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1228..1245
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1252..1962
                     /codon_start=1
                     /product="orange fluorescent protein KillerOrange, a 
                     genetically-encoded photosensitizer derived from KillerRed 
                     (Sarkisyan et al., 2015)"
                     /label=KillerOrange
                     /note="mammalian codon-optimized"
                     /translation="ECGPALFQSDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFNVHA
                     VCETGKLPMSWKPICHLIQWGEPFFARYPDGISHFAQECFPEGLSIDRTVRFENDGTMT
                     SHHTYELSDTCVVSRITVNCDGFQPDGPIMRDQLVDILPSETHMFPHGPNAVRQLAFIG
                     FTTADGGLMMGHLDSKMTFNGSRAIEIPGPHFVTIITKQMRDTSDKRDHVCQREVAHAH
                     SVPRITSAIGSDQD"
     terminator      1984..2078
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2122..2781
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2188..2207)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2846..2932
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2979..3001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3087..3227
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3142..3161
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3242..3259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3413..4001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3493..3512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
       61 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
      121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
      181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
      241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
      301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
      361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
      421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
      601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
      661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
      721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
      781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
      841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
      901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
      961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     1021 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     1081 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgaga
     1141 aatcataaaa aatttatttg ctttgtgagc ggataacaat ttcacacaga attcattaaa
     1201 gaggagaaat taactatgag aggatcgcat caccatcacc atcacggatc cgagtgcggc
     1261 cccgccctgt tccagagcga catgaccttc aaaatcttca tcgacggcga ggtgaacggc
     1321 cagaagttca ccatcgtggc cgacggcagc agcaagttcc cccacggcga cttcaacgtg
     1381 cacgccgtgt gcgagaccgg caagctgccc atgagctgga agcccatctg ccacctgatc
     1441 cagtggggcg agcccttctt cgcccgctac cccgacggca tcagccattt cgcccaggag
     1501 tgcttccccg agggcctgag catcgaccgc accgtgcgct tcgagaacga cggcaccatg
     1561 accagccacc acacctacga gctgagcgac acctgcgtgg tgagccgcat caccgtgaac
     1621 tgcgacggct tccagcccga cggccccatc atgcgcgacc agctggtgga catcctgccc
     1681 agcgagaccc acatgttccc ccacggcccc aacgccgtgc gccagctggc cttcatcggc
     1741 ttcaccaccg ccgacggcgg cctgatgatg ggccacctcg acagcaagat gaccttcaac
     1801 ggcagccgcg ccatcgagat ccccggccca cacttcgtga ccatcatcac caagcagatg
     1861 agggacacca gcgacaagcg cgaccacgtg tgccagcgcg aggtggccca cgcccacagc
     1921 gtgccccgca tcaccagcgc catcggtagc gaccaggatt gaaagcttaa ttagctgagc
     1981 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     2041 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagctagctt ggcgagattt
     2101 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     2161 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     2221 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     2281 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt
     2341 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     2401 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     2461 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     2521 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     2581 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     2641 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     2701 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     2761 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggg
     2821 gtaatgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2881 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccctc
     2941 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3001 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3061 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     3121 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     3181 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3841 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4081 tatcaaaaag gatcttcacc taga
//
