LOCUS       sequence-405174-        4736 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 mCherry gene
            with NLS sequence positioned in between..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4736)
  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 4736)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..4736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1035..1739
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELY"
     primer_bind     complement(1180..1198)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1448..1467
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2821..2838)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2992..3580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3072..3091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3751..4542,4543..4611))
                     /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     4374..4393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4612..4716)
                     /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 aggaggataa catggccatc atcaaggagt
     1081 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1141 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1201 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1261 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1321 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1381 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     1441 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     1501 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     1561 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     1621 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     1681 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtact
     1741 ctagataaac gcggtcaccc gtttctcttc tgttggttac acaatcttgt cacacttttg
     1801 tcatcttttc tgcagctctt tctgccggcg aaacaatatt agaacaagct gttttgcgac
     1861 agtcatctct tgttacctaa gcttaatttt tttgcaaaaa tcaagctcgc cttggatcga
     1921 gtgggaagtt tcatgtagta cataattatt actgtgtaat tatacgggga ttaagttgcc
     1981 attactgtgc ttctttttta atttgtcgag aaagtaagat ggacacctgc tcatttcatc
     2041 catgcgcctt gtttgaacca gcattaagtt aaactacgtt ccttgcatca gacggggttt
     2101 catacagtcg cgcataaccg ccaatgaatt cacgggaaca accaaccgtt cagacggaca
     2161 ctagtgagct cttccgcccc attccggctc ctcggtccag aataaatcag tcgcgaagga
     2221 ttttacacgt cacggaaatc tacaagtgtt ttatctggta cactttaaac cattgcctaa
     2281 aacggcgttc acaagacaag aaccaacgag tacataacat tacatttcgc gctgcagcag
     2341 tttgccatca catggtgtaa tgataacaaa atatggtctg agcgggatta tccataaaac
     2401 tagaagtcag agcttgaaag caagcatcta actgaaatac gttaatcgcg gaaaactgca
     2461 gcaaacgtgt cggatgattt gtaaaagtcc ctgaaatatc ctcgagttcc tataagaaac
     2521 cccgtcaggt ctacgtctcc ttcgtttaac ctcatttcca gatcccgcat tcccggaacg
     2581 tttttcggcc attggtcgac aactaaaaga aaacacatca gtgaaccttc tgtgactaaa
     2641 ataaattcga gaaaaaaatg agttcgtttg aagggtgaaa gagccaacca acgtttgttt
     2701 gattctttct cccccatctc gacaatgact ccagcttctg ctcccggggg atccgctgca
     2761 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2821 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2881 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3421 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgc
//
