LOCUS       sequence-405254-        4764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the tubulin 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 4764)
  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 4764)
  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: pRvTUB-mEGFP-NLS
FEATURES             Location/Qualifiers
     source          1..4764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1036..1038,1039..1041,1042..1554)
                     /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(1081..1102)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1342..1361)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1689..1710
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1753..1773
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2849..2866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3020..3608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3100..3119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3779..4570,4571..4639))
                     /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     4402..4421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4640..4744)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgggtc gatcgacatt gattattgac tagtcaaagc gtcgagggga aagcagcttc
       61 agtgacctga tactggtagc aatcccagac gacggccagc agtgcatttc tagcacaaaa
      121 catcatggtg aatatgttct cacgagaaca tcacggttca gaaatatgct cgtaaacaag
      181 gacatgcaga attacgcaag tggaacttgc cacatcactt cgccgccacg tgcgatgcac
      241 gatctttata ttcaactcac ctaatactgc tctcagcacc agcgtgaaac ccaaacaaga
      301 acccgcccgt cttcttataa aaagtcttcc ggaattttca gcatcacgat gtcctcgctc
      361 cccttcatac tccccgcagc ttccggttag acctacaata tacaagtact gagttaagag
      421 gagacgatgc agacgtctag gacagtaaaa ccgtccgata gcatcgcttg caagaacccg
      481 cagttctttc gacagattcg gaagtccgaa gaggaccttc ttgttttctc acgagaaaga
      541 aaaggtctcc gctacatacg aaccggaaac gcatatatgg acaattttaa acttcaggga
      601 aattgcgatt tctttgacca ggtggacact tcggacttta gagtttagtt taagacgatg
      661 tcaaccacac ttccatcaga gagtatagat tgggctatac tctctgcttc cactgtcgtc
      721 ggtggagtgg aatatagact ggaggccggc ccaccaaggc tggcctgtgg tcggtcaggg
      781 ccaatctaag atacgccgac cactcgatga ctcaactgtg agtaatacac cgaatgacgc
      841 atacgggtgt ttcctatctg actggctaag tgcaaacttt ggcctagaca gcagtcagtg
      901 tgccatttct gccgtagctc ggacgttgaa agatccttct ttcttgcttt tgccgcgttg
      961 tcgacaaccc ctcgacatct tcgtaacgac cttgtggtag ttctccgttg aaggacatca
     1021 tcccccgcat caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1081 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1141 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1201 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1261 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1321 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1381 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1441 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1501 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1561 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1621 ctgcccgaca accactacct gagcacccag tccaagctta gcaaagaccc caacgagaag
     1681 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1741 gagctgtcta gaccaaagaa aaagagaaaa gtataagtat ggatggacat gacaaacaag
     1801 cactctcttc ttccaccaca gaactatcca ggctgctcat tttttattgt gtcgtgtgct
     1861 tctgaccaag ttttaagctc agtctgtcat cggcccatct ttttcttcgt atcttgtact
     1921 ttttttcagc acagggtagc cgcgatattt atggttttat ctacatagat tgtgccgtgt
     1981 aaacttctgt cctggactga atagactgaa taaaatgtct tgaaaatgca agttctgtac
     2041 tttacagaag aggaacgtga gacttcccga aatctagtac tgcacttccg cgcaaacgta
     2101 tcctggccat ttctgctccc agaagcccgc tacacacgac gactgcttca tggtctccta
     2161 agagcctgta acccgggaga aatatcagtc taagcagcta ccgaaacgtt ccctgctata
     2221 gtttatcccg ttttacactt tcccttacac gggtatgtat gtcaccgtga tcgcacgagt
     2281 gacgtctttc atgtcctcgc tagcaggcag tacgccttcg gtttcggaca acccaaacat
     2341 cgaaggacct ccatctaaaa cagagactga tgaacaggag ggacgaacta ccgcgcgacc
     2401 atgatatatc cttcgaaaat cttaccaaag tggagcttcc aagggcaggc tgatttggtg
     2461 ccatttgaca ggtaaaacac tcgttcgtag ggttgctgta ccctcgatac agtaaagtca
     2521 atctctgttc tcgataattt gacgaatggg aggacgatcg tcgctcgaag ggcaacaaac
     2581 tatagagcac agagaagcag acgtaggata accttgaagc gtctggaaga gtagtagcgt
     2641 actcacctgt ggttcttcca gtgagctatt ccatataccc aggtcggacc gaatctccag
     2701 attctgatat cgattttttt caacatgctt tcgaagcacc gtcgcgaact gggaggtcgg
     2761 catgtcgaaa gtcagtggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2821 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2881 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2941 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3001 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3061 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3121 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3181 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3241 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3301 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3361 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3421 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3481 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3541 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4381 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4741 cgcgcacatt tccccgaaaa gtgc
//
