LOCUS       sequence-405259-        4766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The vector contains 1kbp of the upstream and downstream regions of 
            the chas gene (BV898_02951, CAHS 86272) 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 4766)
  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 4766)
  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: pHeCAHS-mEGFP-NLS
FEATURES             Location/Qualifiers
     source          1..4766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1038..1040,1041..1043,1044..1556)
                     /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(1083..1104)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1344..1363)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1691..1712
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1755..1775
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2851..2868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3022..3610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3102..3121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3781..4572,4573..4641))
                     /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     4404..4423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4642..4746)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgggtc gatcgacatt gattattgac tagttatatt taatggtttt aaataaacat
       61 tggcctttaa atattgattt ctatttttgt aacttttgat aaattagcag tatattttat
      121 ttgaaataaa aagtcccttt ttgagccatt caattcaaaa tgtctgttaa acttatatca
      181 acgaaactat taatacatcc aagtcgccac tcgtcatcgt cgttctggtt ttgattacga
      241 accggctcgg tattctacat cacatacagt actgcacggt cgcgaccagt tggtgctgat
      301 tttcaaatcc acccgtatat tatgactgca catactgtgc ttcactacag tcataatata
      361 cggtggatct aaaaagcagc accaattggt cgcgacagta cagtatagta aatttacgta
      421 actagggtgt tactgcattt ccgccccttg acgtcttaga aactaccaat cccatcaata
      481 acaacgaatt tgctttccct tatgtcgtag gatttgtccc atagctggac aaagtcaagc
      541 gaaaaaccta ctcacacccc ggcacaaagt ccagtgaatt tatactactt gcataaattg
      601 tcactgcgca atgtccatcc ctcatcatca tgtctgtgcc tctgcggaaa tcctgtactg
      661 tacattccat ggaattttat cgggaaagag cggtaaatac cgttgcagga ataaccggtt
      721 gatgcgtgga ctaaaattag ccaatccgaa gctccggtat gactcatctg tcgtatttct
      781 accaatcagc actgcacaat gccattaaat cggcccccga caggcgtggc ttggattttc
      841 cgctggggtt tttccactcg tggaatttcg cacgttaact tggaaatttc gcctacatat
      901 agagccgcgc tggacagtct ctcggcatca ttctaacttt ctcgcactga aaatttcttt
      961 gcaaagttcg tgtcccgata ttgattattt tgttcccgaa ggaagtatac agaaccgcaa
     1021 actcacacca cagccccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1081 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1141 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1201 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1261 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1321 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1381 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1441 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1501 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1561 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1621 tgctgcccga caaccactac ctgagcaccc agtccaagct tagcaaagac cccaacgaga
     1681 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1741 acgagctgtc tagaccaaag aaaaagagaa aagtataaga actgatcgct cacccttttt
     1801 aaccagcgac cagaccaacg gatccagttt tcggcgtagc ttatttggtg tctattatga
     1861 aaaaggtggc tgctaaagca gcttcaagct atcctttggt ttccctagaa attcctcctt
     1921 tggtctgtga gactttctga tcttttctcg tattcacgta ctgtgctagt aagttttttt
     1981 agcagcagaa ctgcttctaa tttttctaga ttctccgaag agaatatgat gaaacaaaaa
     2041 gcgtttgtcc tgtccttcct tgtcatcggt gtagacgttg agaaccaaag aagatacaac
     2101 gaccggatga cttcttggaa taatcctcac gcttgcagca ctctactttt ggtaccgtat
     2161 gctcgtgatc tcgagggaca cttttctgcc ctagccgtaa atgttaacca acccgggccg
     2221 tgcgtgtaca gtgccgttca ccaaattaga gagttttcgc aaacggtata tcgcacggta
     2281 tagcggtaca gtatagaaaa ttctctctga ccccagttca tttttttata ctgtaccgct
     2341 atgccgtgcg atataccgtt tgcgaaaact ctctattacc actgtaggga gaaatacacg
     2401 gaaaagcacg actgtacaat caatcaatcg agcacaacga gtacgaaatc agccaataag
     2461 agaagagtga atcagattga acgttcaatc gagagcaggg ccataccgag ggaggcgggg
     2521 cttagctgtc acttgagacc gcgagcatac ggtagttaac acatttttcg tagaatacag
     2581 tgttccaaat cctgggtatc ccatgtgttt aaattaaata ttcgtaacga aaaatgcgtc
     2641 caaaagttat aacaaataaa ctatacagcc gaagatagca aaacaaacgg actctgagag
     2701 gtcaaagctc aaatttatgg aatacgatcg aaacgaaata aatggtgcat tattagtcag
     2761 agttagaaca gaaagatggg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     2821 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3901 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4621 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4741 tccgcgcaca tttccccgaa aagtgc
//
