LOCUS       sequence-405253-        4765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the ubiquitin c gene from Ramazzottius varieornatus, with the mEGFP 
            gene with NLS sequence positioned in between..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4765)
  AUTHORS   Tanaka S, Aoki K, Arakawa K
  TITLE     In vivo expression vector derived from anhydrobiotic tardigrade 
            genome enables live imaging in Eutardigrada.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Jan 31;120(5):e2216739120. doi: 
            10.1073/pnas.2216739120. Epub 2023 Jan 24.
  PUBMED    36693101
REFERENCE   2  (bases 1 to 4765)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-01-31"; volume: "120(5):e2216739120. 
            doi"; pages: " 10.1073/pnas.2216739120. Epub 2023 Jan 24"
COMMENT     Sequence Label: pRvUBC-mEGFP-NLS
FEATURES             Location/Qualifiers
     source          1..4765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1037..1039,1040..1042,1043..1555)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(1082..1103)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1343..1362)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1690..1711
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1754..1774
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2850..2867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3021..3609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3101..3120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3780..4571,4572..4640))
                     /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     4403..4422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4641..4745)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgggtc gatcgacatt gattattgac tagtatccag tcacaacctg ctcccacgta
       61 tactggaaga ccgcggtgct cctgtgcagt cgccccgact ggggttccgg tgaaccatcc
      121 ttgcgaaaaa accaaccgat agtcattgct ccttactgtc ttgctatctt cgaagtgaca
      181 tttcttgcat cacctgccag aaaatgtagt aggttaagac agcagagcca aacgattagc
      241 aaactgcgca acacgactta catcaccggt tcgtccttct tccgctgtcc ccaacgtcgt
      301 atcgttgtcg tgctccccac tacttctgta tgagtactgc gagacttctg tctctccacg
      361 gacagtccaa gtgagacccc ggaatcgacc tgaacgctgg gaacaaaact ggaaaatatt
      421 acaggaactc ggaacttcgg aagggagtca caggacaaaa aatcagctga ttagcgcctg
      481 tctgcaaagt caaacaatca agtcgagtgg gcgacaacac aggcagcagt ctggtggcta
      541 tcgctctgct ggtgatcatc cagcttctac acgcggcgaa cgggtacata cgcaatcaaa
      601 tcaaattcga caatcgacaa tgatcaattt gatatcttga ccaacctacc accacaaggt
      661 taccagtggc agacggaaac ggaacgtgac gtctgactca tccgggcaca gaacaaaagg
      721 cgttcgtcct tcgcctggtg taattttcct ggtaggccga cccagatgac aagcacagtt
      781 cttgactggc tacgtgttcc gtccgtaacc aatgcggtca tctggcttat tcttggcaac
      841 ggaacgagga cgaatgagat cattggaacg ttccagaagg gctttgcccg ggtataaaag
      901 ccgacgtaac gcgctgctag agtttcattg ctcagctttt ctctctcgtt tacccaccac
      961 agcagccatc tttgctcttt gtgtaatttc cctgaccttt tccccgttct agattcctca
     1021 ccacgtctcc cacgccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1081 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1141 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1201 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1261 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1321 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1381 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1441 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1501 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1561 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1621 gctgcccgac aaccactacc tgagcaccca gtccaagctt agcaaagacc ccaacgagaa
     1681 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1741 cgagctgtct agaccaaaga aaaagagaaa agtataagtt gattgactga ttggtccgtt
     1801 ctgatggtct agctttctct ttagttttta cacagcgtcg cttttacaga ctgtatcatc
     1861 tgaactcttt tgttgcgata cgattgtagt cttctgtagt aattccttcg ctctatggtt
     1921 tacgaagaaa aagcgcgtga aagagatgaa atattaaaat gaaaagaagg gaaactttat
     1981 cgggttccta tagaaccaga ataagaaatg atcgagttcc tgttgattgg ttgatctttc
     2041 acactgcggg ctgatcgacc acaagcctac tttgttgttt gcggacagca gcagacaggc
     2101 agccacctcg aatctgacga agttgggcaa tggctgattt atcccatttc ccgctgtaca
     2161 agcgagagct ccggagaaag gtctcgcgcc atcgttggat acagatgaaa ctctaaggtg
     2221 cggaaattta cgagaaggat taacatggaa gtaaaaaccg tcgataagaa gttaactctt
     2281 tgtgtcacat gaagtgcgtc acgacttgat tacgttcaag gggaaatcga tagaaactgg
     2341 cagatactgg ttatcaactg cgttgtgaag tgtacgactc catgggagga catgagcaaa
     2401 caaggttttg gtctccatag aggtcgtcta ccctggcgca tccaggccag actttgttgg
     2461 acgcattgat tctgctggtg gggaaggcag ccttggtgat ggagtagggc aacttccagc
     2521 tgcttccgtc agctccgatg tccttcatag tgtgtggtgc catctttaag gaattcatca
     2581 ggcgtgtcat accttctgga gttgcctctt cgggacccaa cgactctaga tccgtaatct
     2641 ctttcctgat atctacacca aaacacaaat tgacacgcta aaatcctacg gcaaaacaca
     2701 caaatgtgtc tccttggaac atgtcgtcaa cttacgaatg agtgcatcgc agaatctgtc
     2761 catttcttcc agggtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     2821 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2881 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2941 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3001 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3061 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3181 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3421 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3601 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3661 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3721 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3781 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3841 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3901 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3961 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4021 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4081 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4141 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4201 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4261 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4321 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4381 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4441 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4501 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4561 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4621 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4681 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4741 ccgcgcacat ttccccgaaa agtgc
//
