LOCUS       sequence-174484-        5427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express GFP-tagged CAHS1 in mammalian cells transiently.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5427)
  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 5427)
  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-CAHS1
FEATURES             Location/Qualifiers
     source          1..5427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1374..2093
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2215..2336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2252..2271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2306..2325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2343..2798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2480..2501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2692..2711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2825..2929
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2931..3288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2931..3122
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2954..2974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3139..3274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3323..4117
                     /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             3323..4117
                     /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(3377..3396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3987..4006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4305..4324)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4349..4396
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4725..5313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5214..5233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc atgccttacg
      661 aaaagcacgt tgaacagacg gtggtggaaa aaactgagca gcctggacac tcgtcgacgc
      721 accatgctcc cgcccaaagg accgtagctc gcgagcagga ggaagttgtc cacaaagagt
      781 ttacccatac cgacattcga gttccccaca tcgacgcacc tcctccaatc atcgctgcca
      841 gcgcagtggg cttggccgag gagatcgtca gtcatggttt ccaagcctcg gcggcccgca
      901 tcagcggtgc ttccaccgag gtcgatatgc gcccaagtcc caagctagcc gaggaagctc
      961 gtcgtgatgc cgaacgatat caaaaggagc acgagatgat caacagacaa gccgaagcca
     1021 cgctgcaaaa gaaggcggag gagtaccgtc accagactga ggcggaagcc gagaagattc
     1081 gccgcgaact ggaaaagcag catgaacggg acatccagtt caggaaggac ctcatcgatc
     1141 agaccatcga aaagcagaag cgcgaagttg atctggaagc caagatggcc aaacgcgagc
     1201 tggatcgcga agcgcagttg gctaaggaag ccctggaacg ttctcgaatg gccaccaacg
     1261 tggaagtcac gctggatacc gcagcgggac atacagtaag tgggggaact accgtctcca
     1321 gcgtcgacaa agttgaaacc gtccgcgagc gcaaacatca tgatccaccg gtcatggtga
     1381 gcaagggcgc cgagctgttc accggcatcg tgcccatcct gatcgagctg aatggcgatg
     1441 tgaatggcca caagttcagc gtgagcggcg agggcgaggg cgatgccacc tacggcaagc
     1501 tgaccctgaa gttcatctgc accaccggca agctgcctgt gccctggccc accctggtga
     1561 ccaccctgag ctacggcgtg cagtgcttct cacgctaccc cgatcacatg aagcagcacg
     1621 acttcttcaa gagcgccatg cctgagggct acatccagga gcgcaccatc ttcttcgagg
     1681 atgacggcaa ctacaagtcg cgcgccgagg tgaagttcga gggcgatacc ctggtgaatc
     1741 gcatcgagct gaccggcacc gatttcaagg aggatggcaa catcctgggc aataagatgg
     1801 agtacaacta caacgcccac aatgtgtaca tcatgaccga caaggccaag aatggcatca
     1861 aggtgaactt caagatccgc cacaacatcg aggatggcag cgtgcagctg gccgaccact
     1921 accagcagaa tacccccatc ggcgatggcc ctgtgctgct gcccgataac cactacctgt
     1981 ccacccagag cgccctgtcc aaggacccca acgagaagcg cgatcacatg atctacttcg
     2041 gcttcgtgac cgccgccgcc atcacccacg gcatggatga gctgtacaag tgagcggccg
     2101 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2161 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2221 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2281 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2341 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2401 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2461 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2521 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2581 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2641 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2701 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2761 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2821 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2941 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3061 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3121 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3181 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3241 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3301 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3361 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3421 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3481 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3541 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3601 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3661 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3721 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3781 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3841 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3901 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3961 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4021 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4081 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4141 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4201 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4261 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4321 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4381 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4441 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4501 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4561 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4621 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4681 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4861 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5281 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5341 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5401 gtggataacc gtattaccgc catgcat
//
