LOCUS       sequence-405248-        5268 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 
            RvSAHS1-mEGFP gene positioned in between..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  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 5268)
  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-SAHS1-mEGFP
FEATURES             Location/Qualifiers
     source          1..5268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1561..1563,1564..1566,1567..2280)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELSR"
     primer_bind     complement(1606..1627)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1867..1886)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2214..2235
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3353..3370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3524..4112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3604..3623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4283..5074,5075..5143))
                     /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     4906..4925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5144..5248)
                     /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 agcccgaatt cgccatgtct cgcgcagctg tcgctatcgc ccttctgggt
     1081 tgcgttgtgg ccgcttacgg cgctccggct gaaggccacg acgatgccaa ggcagaatgg
     1141 accggaaaga gctggatggg caagtgggaa tccactgatc gcatagagaa ctttgacgcc
     1201 ttcatctccg cccttggtct tcctctcgaa cagtacggtg gaaaccacaa gaccttccac
     1261 aagatctgga aggagggtga ccactaccac caccaaatca gcgtccccga caagaactac
     1321 aagaacgacg ttaacttcaa actcaacgag gaaggaacaa cccaacacaa caacacggag
     1381 atcaagtaca agtacaccga ggatggcgga aacctgaagg ctgaagtcca cgttccatcc
     1441 cgaaacaagg ttatccatga cgaatacaaa gtcaatggag acgaactcga gaagacctac
     1501 aaggttggag atgtcaccgc caagagatgg tacaagaaga gctcttcgtc gggcggccgc
     1561 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1621 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1681 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1741 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1801 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1861 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1921 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1981 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2041 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2101 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2161 tacctgagca cccagtccaa gcttagcaaa gaccccaacg agaagcgcga tcacatggtc
     2221 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtctagataa
     2281 atgtttgacg aatgccatat aactgttcca tttttcgttc tgatgtaaac actttttcgc
     2341 tgtcgcattt atccatctgt agctagaaat cttgacagct ctttcctcat tcagtttccc
     2401 tatgcaagtt cgacctgctt ttggcacgaa ctcttacatt gaaaccgctc atcgaaacct
     2461 ccactcactc tctcattatt cggcacgctt tggtcccaag tcccaagttt cgatcctacc
     2521 ccaagatgag gcaagtgaga tccatgatgg cgtaaaggtt tggtacaggc taagccgaaa
     2581 acaaatattt cgcgaatcag accatcaaac caccatgcat gcgccgcgcg gctgcctcga
     2641 aggtgacact gagtggccaa agctgatcaa gctgtcgttg aaaaaaggaa tcagctagtt
     2701 tcatcccgat gtcgataaaa tgaaacaggc gcagaatgtt taagaccatg cttgagggac
     2761 gaagggttat aggcgagtcc atgaaaatca aacggtgtaa ggtaaagatt cgctgaggca
     2821 ccaggacgta tacagcgtat cttgtgttta aatgaaatca aattcgccgc taacttaaat
     2881 gacgagcgtt ttctggacta cgctcagggt ctccaaatcg aagcctaaat ttgagcgtgt
     2941 agtctgcaaa aatactgatt cttcttagga agtcgttccg agaggttttg tgctgggcga
     3001 gtagccgtag cctgctatat cacagagaac cttctggaaa atcttctcgg ggtcgtccgt
     3061 tccttcgata taacctgtca taaacaatga tccaaatgta gcagcatcta gggagtacca
     3121 ccttttgtaa taagctttac agaaccgtac catcgcgaca aagaggtttt ccttacttac
     3181 agatatcttt gtatcttctg tgaagttgag cggaataata ctcatgcatc ggataaacga
     3241 cttgctcgta atatcccggc acatccggtg ggtcgtagtt ggatccgctg cattaatgaa
     3301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3961 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgc
//
