LOCUS       sequence-405250-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the cahs3 gene from Ramazzottius varieornatus, with the mEGFP gene 
            positioned in between..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  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 4743)
  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: pRvCAHS3-mEGFP
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             911..925
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             join(1036..1038,1039..1041,1042..1755)
                     /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(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"
     primer_bind     complement(2828..2845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2999..3587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3079..3098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3758..4549,4550..4618))
                     /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     4381..4400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4619..4723)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgggtc gatcgacatt gattattgac tagttgttct cgggaatcca ccccctcgat
       61 ttctacgaat tcttctttgt aaaatcccca aggaccatct tcttgctttc ttttccggta
      121 ctgatagctc aaaggtgctg ctgcatagtg gccgtgttgc cggtgcgatg gcggagctga
      181 ccgtgtgata aaaaaatcgt aaaataatcc tggttaaaca tgcatcacta ttattctaaa
      241 actcttcctt cttcccgctt tttgtgtttc aagtggatct tgagcaaaaa aaacaaaaga
      301 ctttgttact ggtgctggac atcagccaag cttcaaaagt cgcgttactt acatacgaaa
      361 ttcacaattt tgacgcaaaa aacggcaaat ttccactgaa agacctatat aataacgtcc
      421 acttcgttga attgtttgct gacgttctta gtgttcgaag agaatatttc ggtggaatgc
      481 ggttttcttt ggcaatatgc gcgaaaaacc tttattctgt ttgctattcg cacctatcga
      541 tagtgaacga cacatctctt gtaaacatcg ccttcaacct atacgggctg ttattcttct
      601 tcttaagtaa tgagtcaagc tttggaaata ccagacggct cgccgctcga gtcgacaagc
      661 atattctcat gacatattgc cgtctgtacg acaggccact cgcattcttt tccaggcgta
      721 cagccggttt ttgccgtgga ttaactctca taggacctct cacgctggtg atatcaggaa
      781 atcacaccac acaagctgtt gttatcgcgc tttgatgatc actacctgta caactcgcag
      841 cattttccgt cgggaaaaag cctggcagaa agttcagctg agcacacaag acgacgacga
      901 cgacgacgac gacgacgacg ataaaacttg ccgactgtgt cagttcagct tcacactatt
      961 cccgtgtact ttgcagcact tacccagctc ttgtttattc tttcgcagcc tttcgctaac
     1021 gccctgtcgc aaaccatggt 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 gataaatttg ctcataacca caccatcata caatttttca ccatttctaa
     1801 aaagaaaatg ttttcgtcca gtcttttgtg cactctgaaa tgtcatatca tgaataactt
     1861 cagttttagc gccgcaagga aaatgtgaga cggttaatat cacgaacgcc ggtaaacaat
     1921 attcgtggtc aataaaacga ttaatttcta catcagtcat gctttcttcc gtgcactgct
     1981 ttaaacaagg caaaccgatt tctaacggaa gaatgtacta ttacctctgc tgaactgtgg
     2041 cccccacgac ttttctttgt ggaatttacc gtaaacgttg agtgcggtac tcaatattca
     2101 agaaaatact ccatgcgtgc acatattacg taagaatgac ttctcgacca ttgtctaaaa
     2161 gtcactggcg catatgtgca gtaccctgac cattgcaaga caggcagatt gcatgctgtg
     2221 ctggtgatcg cccagggctg ttttttgttt actgtcacag accgcgactg ccttcgcaat
     2281 gactgatccc tcccaccgaa gtaaacataa ttatgaccac aaaatcgagt tttattacat
     2341 gtcacaaaag tggctgcaga agatctcaca gattatgaca actcatcgta aaattattat
     2401 caaagttcaa attcgcggag caaaatggct caccagataa ctatgctaag atcagacttg
     2461 gtagcatact cagtggatat gtcttgtcaa agtggggtcc acggattccg catttgtcac
     2521 agtagttgtg cgtgtggtgg tcacaccagt ggtcagatcg ctgcctacgc cccctgtagt
     2581 cgtcccttga gtcatcccaa ggactctttc atccggaacc aaatcggctt gacgcttgtc
     2641 tttgaaattc atgccggggt cctcgccacc ttggtcttcg cggacatcta cagaaacgac
     2701 gtttccgtct ccgtcgacat gcgccttctc cacccgctca tactttacct cctttggatc
     2761 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4741 tgc
//
