LOCUS       sequence-393242-        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 355, Emission: 424.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  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 4732)
  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-N1
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             154..948
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             154..948
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(208..227)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     818..837
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1136..1155)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1180..1227
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1556..2144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2045..2064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2319..2622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2623..2826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2777..2797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2849..2929
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2931..2940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2982..3003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3243..3262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             3402..3653
                     /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"
     polyA_signal    3778..3899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3815..3834)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3869..3888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3906..4361)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4043..4064)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4255..4274
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4388..4492
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4494..4732,1..119)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4494..4685
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4517..4537)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4702..4732,1..105)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
       61 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
      121 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      181 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      241 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      301 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
      361 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      421 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      481 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      541 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      601 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
      661 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
      721 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
      781 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
      841 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
      901 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
      961 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     1021 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     1081 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     1141 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     1201 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     1261 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     1321 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     1381 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     1441 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1501 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1561 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1621 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1681 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1741 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1801 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1861 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1921 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1981 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2041 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2101 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2161 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2221 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     2281 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2341 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2401 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2461 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2521 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2581 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2641 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2701 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2761 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2821 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca
     2881 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg
     2941 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3001 acgtaaacgg ccacaggttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3061 agctgaccct gaagctcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3121 tgaccaccct gcaattcggc gtgctgtgct tcgcccgcta ccccgaccac atgaagcagc
     3181 acgacttctt caagtccgcc atgcccgatg gctacgtcca ggagcgtacc atcttcttca
     3241 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3301 accgcatcga gctgaagggc ttcgacttca aggaggacgg caacatcctg gggcacaagc
     3361 tggagtacaa cgggataagc tcaaacgtat atatcaccgc cgacaagcag aagaacggca
     3421 tcaaggccca cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     3481 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3541 tgagcgtcca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3601 tggagtccgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagcgg
     3661 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3721 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3781 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3841 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3901 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3961 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4021 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4081 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4141 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4201 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4261 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4321 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     4381 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     4501 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4561 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4621 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4681 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc cc
//
