LOCUS       sequence-405318-        6141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Sirius, a pH-insensitive and photostable 
            ultramarine fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6141)
  AUTHORS   Tomosugi W, Matsuda T, Tani T, Nemoto T, Kotera I, Saito K, Horikawa
            K, Nagai T
  TITLE     An ultramarine fluorescent protein with increased photostability and
            pH insensitivity.
  JOURNAL   Nat Methods. 2009 May;6(5):351-3. doi: 10.1038/nmeth.1317. Epub 2009
            Apr 6.
  PUBMED    19349978
REFERENCE   2  (bases 1 to 6141)
  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.1317"; journalName: "Nat Methods"; date: "2009-05"; 
            volume: "6"; issue: "5"; pages: "351-3"
COMMENT     Sequence Label: Sirius/pcDNA3
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        201..404
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     355..375
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     449..468
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        449..467
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(545..566)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(806..825)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             965..1216
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKAHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSVQSKLSKDPNEKRDHMVLLESVTAAGITLGMDELYK"
     promoter        complement(1280..1298)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1281..1298)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1318..1335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1324..1548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1594..2022
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1681..1700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1891..1912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2031..2051)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2036..2365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2216..2351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2278..2297
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2432..3226
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKYRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2486..2505)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3096..3115
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3400..3521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3437..3456)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3491..3510
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3570..3586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3570..3586)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3583..3605)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3594..3610
                     /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)."
     promoter        complement(join(3618..3624,3625..3642,3643..3648))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3663..3684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3801..3818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3972..4560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4052..4071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4731..5522,5523..5591))
                     /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     5354..5373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5592..5696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5771..5790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5962..6141,1..200)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
       61 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      121 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      181 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      241 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      301 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      361 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      421 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      481 ggtaccgagc tcggatccga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      541 catcctggtc gagctggacg gcgacgtaaa cggccacagg ttcagcgtgt ccggcgaggg
      601 cgagggcgat gccacctacg gcaagctgac cctgaagctc atctgcacca ccggcaagct
      661 gcccgtgccc tggcccaccc tcgtgaccac cctgcaattc ggcgtgctgt gcttcgcccg
      721 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      781 ccaggagcgt accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      841 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      901 cggcaacatc ctggggcaca agctggagta caacgggata agctcaaacg tatatatcac
      961 cgccgacaag cagaagaacg gcatcaaggc ccacttcaag atccgccaca acatcgagga
     1021 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1081 gctgctgccc gacaaccact acctgagcgt ccagtccaag ctgagcaaag accccaacga
     1141 gaagcgcgat cacatggtcc tgctggagtc cgtgaccgcc gccgggatca ctctcggcat
     1201 ggacgagctg tacaagtaag aattctgcag atatccatca cactggcggc cgctcgagca
     1261 tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     1321 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1381 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1441 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1501 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     1561 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     1621 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1681 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1741 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1801 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1861 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     1921 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1981 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2041 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2101 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2161 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2221 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2281 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     2341 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     2401 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2461 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2521 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2581 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     2641 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2701 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2761 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2821 ctgcccattc gaccaccaag cgaaatatcg catcgagcga gcacgtactc ggatggaagc
     2881 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2941 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3001 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3061 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3121 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3181 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3241 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3301 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3361 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     3421 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3481 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     3541 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3601 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3661 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3721 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3781 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3841 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3901 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3961 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4021 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4081 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4141 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4201 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4261 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4321 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4381 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4441 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4501 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4561 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4621 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4681 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4741 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4801 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4861 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4921 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4981 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5041 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5101 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5161 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5221 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5281 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5341 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5401 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5461 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5521 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5581 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5641 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5701 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     5761 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     5821 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     5881 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     5941 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     6001 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6061 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6121 ccatagtaac gccaataggg a
//
