LOCUS       sequence-405173-        4763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the actin 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 4763)
  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 4763)
  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: pRvACT-mEGFP-NLS
FEATURES             Location/Qualifiers
     source          1..4763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1035..1037,1038..1040,1041..1553)
                     /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(1080..1101)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1341..1360)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1688..1709
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1752..1772
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2848..2865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3019..3607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3099..3118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3778..4569,4570..4638))
                     /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     4401..4420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4639..4743)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgggtc gatcgacatt gattattgac tagtcgacag tcaaccgttt tgaggacaag
       61 agggaaaatt ccagtcgcat cagatcggaa tatcaaggtt gatggttgaa gtccatacgg
      121 tgctgaaggg agttcgaaga gttttgttgt ataatctggt gagggaacat tgttgcgttt
      181 ataggcgaag ggaggaaact tttcacgcat gacatggtcg atgttttgtt ctcaaaattt
      241 gggaatgtct ctttgcatgg agctagtaaa taaattggtt gatcgacatg aacatgctag
      301 caacttgagt ttggatttct ggcacaaaag ttgaagtttt ttattttgag gcattcataa
      361 ttatgttttt gagttccgca agattgttct agctgggatt tattttccat ttcggaataa
      421 acgaccatca ttgccgaggt gagggcggat gaatggagga attcaatatg gcgtgaaggg
      481 cagcaaattt actgaatgcg aagcgtatga tcctgtcaga ggttctgtct cttatgtgcg
      541 aatcagagag gcgaattctg aaagcagacg agagtgtcta gccagctcag ggaagaattt
      601 tgcacttgtt cccgttgtta ttaccgatct ctttgattcc gttcccagaa gaagtccatt
      661 cagagaaatc tccgcgtgtt tcccaaagca cgttgctgaa tttatttttg gctcacgaag
      721 aagcccggcg attacttgaa atggacagat cgtagctagc gtagtcgaga atgcgagagt
      781 gtagcgaaga tggctaaaat tagcgagaga ctgtgtatgg gtacggatat gagtatgttg
      841 atatgaagct gaagctaccc tcgacggaag cgcacaagag ggcgtttact cgctgttggc
      901 catgtatgga ccgaagatta gacatagcaa tgtcgcatgc ctgtcatcat tgaatgtaat
      961 cattgcaaac ctctttactt gattttgttt tctctgttgc agttttcctc ctttttccac
     1021 aaggttaagc taccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1081 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1141 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1201 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1261 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1321 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1381 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1441 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1501 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1561 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1621 tgcccgacaa ccactacctg agcacccagt ccaagcttag caaagacccc aacgagaagc
     1681 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1741 agctgtctag accaaagaaa aagagaaaag tataaacgcg gtcacccgtt tctcttctgt
     1801 tggttacaca atcttgtcac acttttgtca tcttttctgc agctctttct gccggcgaaa
     1861 caatattaga acaagctgtt ttgcgacagt catctcttgt tacctaagct taattttttt
     1921 gcaaaaatca agctcgcctt ggatcgagtg ggaagtttca tgtagtacat aattattact
     1981 gtgtaattat acggggatta agttgccatt actgtgcttc ttttttaatt tgtcgagaaa
     2041 gtaagatgga cacctgctca tttcatccat gcgccttgtt tgaaccagca ttaagttaaa
     2101 ctacgttcct tgcatcagac ggggtttcat acagtcgcgc ataaccgcca atgaattcac
     2161 gggaacaacc aaccgttcag acggacacta gtgagctctt ccgccccatt ccggctcctc
     2221 ggtccagaat aaatcagtcg cgaaggattt tacacgtcac ggaaatctac aagtgtttta
     2281 tctggtacac tttaaaccat tgcctaaaac ggcgttcaca agacaagaac caacgagtac
     2341 ataacattac atttcgcgct gcagcagttt gccatcacat ggtgtaatga taacaaaata
     2401 tggtctgagc gggattatcc ataaaactag aagtcagagc ttgaaagcaa gcatctaact
     2461 gaaatacgtt aatcgcggaa aactgcagca aacgtgtcgg atgatttgta aaagtccctg
     2521 aaatatcctc gagttcctat aagaaacccc gtcaggtcta cgtctccttc gtttaacctc
     2581 atttccagat cccgcattcc cggaacgttt ttcggccatt ggtcgacaac taaaagaaaa
     2641 cacatcagtg aaccttctgt gactaaaata aattcgagaa aaaaatgagt tcgtttgaag
     2701 ggtgaaagag ccaaccaacg tttgtttgat tctttctccc ccatctcgac aatgactcca
     2761 gcttctgctc ccgggggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2821 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2881 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2941 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3001 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3061 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4381 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4741 gcgcacattt ccccgaaaag tgc
//
