LOCUS       sequence-405176-        4762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the actin gene from Hypsibius exemplaris, 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 4762)
  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 4762)
  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: pHeACT-mEGFP-NLS
FEATURES             Location/Qualifiers
     source          1..4762
                     /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(2847..2864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3018..3606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3098..3117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3777..4568,4569..4637))
                     /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     4400..4419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4638..4742)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgggtc gatcgacatt gattattgac tagtacttga ttgttagtaa tttttaaaca
       61 cagaaaaatt tctttgtatg ctacttcgtc ggccatgtcg gagggcgtca aagatgacac
      121 atcgacgacg ctctctcatt cagaaccttg tgtgtatttg tctcccgcta ggtggcggta
      181 tcggtcactg ggcccttgaa tcaggtgcag gcggcgtaag gcgtttctct ctccacgttc
      241 ccaggcgttg ctttgcattt cgaaatatat atcgagcaaa atctccgttt ttcctttaaa
      301 agtttcaaat tctcctcccc cccatttccg ttcattccac cacatcgcta gccaacgcct
      361 ctccagtttt cgggacaaat aaggcatgtg tctgttaata tctgtgtgta tatgtaacac
      421 ttcagcagct gaggaaaggg atatctctct gagggtgtta aaaataacat ttttccccta
      481 tctcaacgca cgacgcgggc catgtgtggt tctggagcgg ttgcgcggat tacttaactt
      541 ggtcatcgct tctgagtgtg tgtgtctgtg aggggagaag atatagcgcg cggaccggag
      601 atgccgaaga gtgagatcta attttggagt gagacagaca gacacgacac acacatagga
      661 tgctgtagcg tggcgtgcac agtgaaaaag ttacactcgc gcttgccggg cgcgcggaag
      721 agggcgtcag aggcggcggt gtactagtcg ctctctcgcc tagtctctca tcgccattta
      781 aggcctgtgt gtgctcttgt gctgtgtgcg tgtgtactag tatgactttg actgtcagtc
      841 tgtgggctca tccatcagtc atccatcctg ccattccaat ccgctggggc ccctctctca
      901 tactctcttt agttcggcat tatcagtttc cgttcatccg gtcaactttt atttatcagt
      961 ttgaagaagc ttttccgcta acgtgtctgt tctctccccg ttgcagtttc gcgtaatccc
     1021 tcagaacagt cgcaccatgg 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 agtataaatt gtcgtcgtcc tgtcctctgt
     1801 cctactactt cttttgtagc gctacgtggt catctcgcag ctctagctgg gatctttttg
     1861 ttgtgtgctt cgacacaaaa aacacacaat ctgtttctgg cgcggtcatc atttttttct
     1921 ctcttgtcgt tacctaagct tattttcaga tccaagctct cctttgagtt ggttacacca
     1981 tgttgataga cacgtgttat tttgcagtcg tcctaaaaat tcgagaatca agcgcgcttt
     2041 tgtagttgcc atttaattat tctattatat aaactgattg ggaaaaacag tgtgcttgta
     2101 tggaattgat taaggtttag acaagtagac aatagtaagt cataaacata aaataattca
     2161 aaaccgagac attaactcag tagtagtatt ggacgatagt cgcgtattga ttcacccacg
     2221 gtctggcgca tggagattgg aaaagctcac gggcctatag tgctgacggt cagataaaat
     2281 ttgtaaacac gaccgtcacc cacgagccaa aagtgttgat gttaggaatc actttttact
     2341 attggcccgt ccttgtatat caaaatttgc ggacgggcca atagtgggaa aacagcaaaa
     2401 gtgcccacgg gcctggccca cacgcttaac gtgtcccggc ccacgggcct ggcccatagg
     2461 ctcaatgcgt ccctgcccac attcaatcat cgatgaaata ccagccggat tgatcagtca
     2521 ccaacaagag tcctgtatta attgcacttt gcttcagcca ggcattaggt tcatcctcgg
     2581 aattttcggc ctggcagcgt agtatttgta ccgcggcaaa taaacgttgg gaagctaaaa
     2641 ttgaaaaaga atagaatggt ttgtccccca cagacatgcc tgagacgttc cgagttcaag
     2701 acggcgaatc gaaatggcga gaaaaaaggc caaagaaata aaaagtggac gctcaatcaa
     2761 actagcgcgt cctaggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2821 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2881 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2941 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3001 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3061 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3121 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3181 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3241 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3301 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3361 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3421 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3481 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3541 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3601 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3661 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3721 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3781 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3841 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3901 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3961 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4021 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4081 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4141 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4201 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4261 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4321 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4381 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4441 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4501 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4621 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4741 cgcacatttc cccgaaaagt gc
//
