LOCUS       sequence-405182-        4763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the sahs1 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: pRvSAHS1-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 tagttaacaa gcagcagaac tgagccctgt
       61 cgcctccatt aattgtgata gttctgcctt cgagttatga ttcctcatgg aagtgctttt
      121 catgtatgtc tgccatttta actaactgtt ggatgttcat ttccatttcg ggatagctct
      181 gggttttttg acagaataac cctgccgcgt ttctgtcttt tcacagtcat cgtttctgct
      241 gtctagaggt tttctgtggg aaactttgcg cggtgacgaa ccatattttt tttgtcccag
      301 tgaatatcat gtactgcgat ctacgacgga agcgctgcct acatgcaaaa aacggaaaat
      361 gttccgcgtg agaactagtt tcctagacat tccgatacaa tcactatcgt caacaacgct
      421 gactggcctg aacggggctg gttagagcct cgccgtgctg taagccagtg aggactcggg
      481 tacgctgtat gaagttagaa aatggctgtt gtttttcagc aaaaggtagt atgacaggca
      541 atcacaaaaa ctcagcggtc ttattgtttt cgaaagtagg tacaaggtac ttttctgaga
      601 agcaagttcg actttttggg acactggcca gaaagaaatt cagacaacac gaggtagaca
      661 acaaccctta gcatggtttt ccaccattca gcaaatgagg ctcaacactt tcagatcacg
      721 cccggaacaa gctcttaagg tctggtttca ctcccgcgaa catcaatggg aagacaaagg
      781 cagttcgccg tggcccaaat aagttcagga aaaagtcttg gtgaaacacc gccaccgcct
      841 tgtccagtca ttctattacc gaaaaattct cgaaagttcg ccactaacct ttgctcttcc
      901 gtgatacata cagctgatta tcgcatcgaa gccccaaact tgaccagtat aaaatgatcg
      961 ccatcgaaaa gacttcctca ttccaagttt tcagtgtttt ctcagcgcat tcctgccata
     1021 caaccaaacc agccatggtg 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 tataaatgtt tgacgaatgc catataactg
     1801 ttccattttt cgttctgatg taaacacttt ttcgctgtcg catttatcca tctgtagcta
     1861 gaaatcttga cagctctttc ctcattcagt ttccctatgc aagttcgacc tgcttttggc
     1921 acgaactctt acattgaaac cgctcatcga aacctccact cactctctca ttattcggca
     1981 cgctttggtc ccaagtccca agtttcgatc ctaccccaag atgaggcaag tgagatccat
     2041 gatggcgtaa aggtttggta caggctaagc cgaaaacaaa tatttcgcga atcagaccat
     2101 caaaccacca tgcatgcgcc gcgcggctgc ctcgaaggtg acactgagtg gccaaagctg
     2161 atcaagctgt cgttgaaaaa aggaatcagc tagtttcatc ccgatgtcga taaaatgaaa
     2221 caggcgcaga atgtttaaga ccatgcttga gggacgaagg gttataggcg agtccatgaa
     2281 aatcaaacgg tgtaaggtaa agattcgctg aggcaccagg acgtatacag cgtatcttgt
     2341 gtttaaatga aatcaaattc gccgctaact taaatgacga gcgttttctg gactacgctc
     2401 agggtctcca aatcgaagcc taaatttgag cgtgtagtct gcaaaaatac tgattcttct
     2461 taggaagtcg ttccgagagg ttttgtgctg ggcgagtagc cgtagcctgc tatatcacag
     2521 agaaccttct ggaaaatctt ctcggggtcg tccgttcctt cgatataacc tgtcataaac
     2581 aatgatccaa atgtagcagc atctagggag taccaccttt tgtaataagc tttacagaac
     2641 cgtaccatcg cgacaaagag gttttcctta cttacagata tctttgtatc ttctgtgaag
     2701 ttgagcggaa taatactcat gcatcggata aacgacttgc tcgtaatatc ccggcacatc
     2761 cggtgggtcg tagttggatc 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
//
