LOCUS       sequence-405257-        4764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the cahs1 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: pRvCAHS1-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 tagttcagaa ctggtccgtc ttgctgagaa
       61 gggtgttgca acttacgacc accggcagcc cacgccactg cgataaaaaa ctattattac
      121 aagactaagc aataaaaaac ccaagtggtc atctgggtca gtatgacagt tatggtgaaa
      181 ggtgcgtaaa agaaacaacc tcgaacgccc atcaaatata gcgtgtcgga tactgaacga
      241 caagaaccgt gggagggttt gcgccttcac gacaagcgtg gtcagcgaac gtggccgaga
      301 acttgacagc acacatcgag gccggaagtt caaggggact agcggtaaga gttctctctt
      361 gagatattaa ccttcatcac tgacactgtt actttttagc gagagcaaaa actacttggt
      421 ccacatttgc actttcgggg ctgaattaat gcgaccaaac catgaagggg tctcctgatt
      481 agtccgcgca aaacaaagaa atcggtaacc atacagcaat ttcacagcca taagtactgt
      541 tcgactcaac acacggagcg tttttgggtt tcaaatcgta aaaaaggcac tgtgaacgcg
      601 gaaggacctt gaggcatcca gtttggccct taaaacggat gattacctca acgatcggaa
      661 tgctacgttg aattccatgt gagacctcgt ggtggttcgc ctcgcaaaat attgccatgc
      721 acatgtgccg ttccgagctc gagacgagac ccgaagtaag acattttgtt tacggttttg
      781 cttacgccgg taagagcgcc gattaaactt ccttgtgaca ctgtctgaag aaaactcgca
      841 acctagtagc ctggagcaaa tcctgcggca acatagactt caccaacttc tggaaagata
      901 gtcaccaaca ggccatggca ttttccgctc atgagttacg agaaaacgga acttgatatc
      961 acaagatagt cattacgttt tctgcttatt cccttttttc gttccagcag taattactcg
     1021 gataatattt cacacatggt 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 gtataacgac cacaccacga ttagaacttg
     1801 cttgcctcat tggatgaacc ctttaattca caaaatatct aagatactca agatcgtaag
     1861 atttagaatg gttagcttct tcatttccgc gtcgacctga atagtacgcg tacggacctc
     1921 gtgaattagt cgcataacat cctaatattt tcaaagtgat aagcgagtct ccaactataa
     1981 acagggcgtc gtaccataat ttccacttga cagtattgct cggcgcgaca actcgagcca
     2041 actgaagcca ctgcgttgca gaactatgtg cgcgacctgt gtatagtgga gttattcaag
     2101 agttttatcg ctaacgactc gttttattta acgaaaagtc acggtgaggt acccgcaatc
     2161 attcattcgg cattccggtg tttacacctt ctgtttgcat tgctcattat atttccgtga
     2221 taagagataa cacccgagct gcatcagcag gttgcacatc ggctggcaaa ttccatgatc
     2281 gcaacataca ctaccgtcac aagctagaaa caccctaaat ttacgtgtca taccgagagt
     2341 tcccttataa ggcaaaaggc ccaggttatt catttcgagc tgtttgtgtt cttgtttatc
     2401 ccaaaattca gccttccttt aaaatggaac gaacagtgga aaaaagcgtt gaagtacgca
     2461 cgaccggaag cagtcatcct tccaacgtaa ccagttctca gtatactgtc gaacgagaga
     2521 caaccatccc agccacacta ggcgatgcaa cctacactgg cacaccttat cctcatggag
     2581 gactcggaca ttcctcatcg cagagttcgt cggacagcca agaacgcgtg accaactaca
     2641 cccacacgga agtgagggtt ccccaggtgt cgatgcccgc ccccatcatc gttacgtccg
     2701 cttcaggact agccgaagag atggtaggtt caggattcac ggcttcggcc tctcgagtca
     2761 ctgggtcgtc ggtggaggat 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
//
