LOCUS       sequence-178996-        5625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express GFP-tagged CAHS3 in mammalian cells transiently.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5625)
  AUTHORS   Yamaguchi A, Tanaka S, Yamaguchi S, Kuwahara H, Takamura C, 
            Imajoh-Ohmi S, Horikawa DD, Toyoda A, Katayama T, Arakawa K, 
            Fujiyama A, Kubo T, Kunieda T
  TITLE     Two novel heat-soluble protein families abundantly expressed in an 
            anhydrobiotic tardigrade.
  JOURNAL   PLoS One. 2012;7(8):e44209.
  PUBMED    22937162
REFERENCE   2  (bases 1 to 5625)
  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: "PLoS One.";
            date: "2012"; volume: "7(8)"; pages: "e44209"
COMMENT     Sequence Label: pAcGFP1-N1-CAHS3
FEATURES             Location/Qualifiers
     source          1..5625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..291
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(293..397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      424..879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(511..530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     721..742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    886..1007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(897..916)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     951..970
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1129..1848)
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     promoter        complement(2852..3055)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2881..2901)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3056..3359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3534..4122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3614..3633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4451..4498
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4523..4542
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4730..5524)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4730..5524)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4841..4860)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5451..5470
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5559..5625,1..291))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5573..5625,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc
      301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      421 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      841 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgccttaaga tacattgatg
      901 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      961 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1021 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1081 acctctacaa atgtggtatg gctgattatg atctagagtc gcggccgctc acttgtacag
     1141 ctcatccatg ccgtgggtga tggcggcggc ggtcacgaag ccgaagtaga tcatgtgatc
     1201 gcgcttctcg ttggggtcct tggacagggc gctctgggtg gacaggtagt ggttatcggg
     1261 cagcagcaca gggccatcgc cgatgggggt attctgctgg tagtggtcgg ccagctgcac
     1321 gctgccatcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccat tcttggcctt
     1381 gtcggtcatg atgtacacat tgtgggcgtt gtagttgtac tccatcttat tgcccaggat
     1441 gttgccatcc tccttgaaat cggtgccggt cagctcgatg cgattcacca gggtatcgcc
     1501 ctcgaacttc acctcggcgc gcgacttgta gttgccgtca tcctcgaaga agatggtgcg
     1561 ctcctggatg tagccctcag gcatggcgct cttgaagaag tcgtgctgct tcatgtgatc
     1621 ggggtagcgt gagaagcact gcacgccgta gctcagggtg gtcaccaggg tgggccaggg
     1681 cacaggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     1741 gccctcgccc tcgccgctca cgctgaactt gtggccattc acatcgccat tcagctcgat
     1801 caggatgggc acgatgccgg tgaacagctc ggcgcccttg ctcaccatga ccggtggatc
     1861 tcggcgttca gtgtgttgtt cggtatgttt aatctgagtg acggtggtgg gtccagtctt
     1921 ggtggtgctg ctcttgtcgg cggaagtctg gattgcaccc ttggagacgg tggtaccaat
     1981 agcggactcg accttgacat tgacgttagt ctgggccttg gcgctttcca aagcctcttt
     2041 agcggcattg cgctcaagct cgagggctcg cattgcgtac tcagcctcca gctggatttc
     2101 tctctcttgt cggttgactt ggttctcaac catttccttg cggaattcaa gatcacggat
     2161 gtgttgtcgc tccaagatct gtcggatgag ttcggcatca cgctccactt cctctcggta
     2221 ctgggcagcg cgcatacccg cgtcttgctc atcttgcctg gccagttgct cgaccttggc
     2281 cttgtagcga gcatactctt caggtcccaa atccttgcgc tggatcttgc tcaagtcaac
     2341 ggtggtccct gcggtaattt cagcgcgagc tccaccgccg aggttaaggt cgaggttgct
     2401 gtggatggga gcagcgatgt tgggagcagc aacggtcaag ttcttgtctt gggtgacggt
     2461 gaagccggtt cggacagatt ccatgttggt gctgccgaaa gttgtcactg ggacggtgac
     2521 gacgtgggtc tcgctttcgg ttttggtacc atgcttgatg acctcacgct cctcatggac
     2581 ctcggagtga gatccagggg ggacggggac agttcgctgg gctccaccga tggtaggtcc
     2641 tccactgctg gtgtggacct cggtgcgcga gtagctggaa cttccttgaa caccttgacc
     2701 accttgacca ccttgctggc tggaggacga gtgttggctg gacgattgct ggttctgtcg
     2761 ggaagacatg gtaccgtcga ctgcagaatt cgaagcttga gctcgagatc tgagtccggt
     2821 agcgctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta
     2881 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     2941 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     3001 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     3061 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     3121 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     3181 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     3241 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     3301 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     3361 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactaa
     3421 tgcatggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3481 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3541 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3601 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3661 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3721 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3781 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3841 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3901 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3961 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4021 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4081 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4141 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4201 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4261 taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg
     4321 agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg
     4381 tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     4441 ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc
     4501 atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg
     4561 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
     4621 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     4681 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     4741 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     4801 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     4861 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     4921 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     4981 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     5041 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     5101 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     5161 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     5221 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     5281 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     5341 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     5401 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     5461 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     5521 tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca
     5581 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcg
//
