LOCUS       sequence-334477-        9189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the full Leptodactylus latrans Na+K+ATPase with ATP1A1-S 
            variant with Q111R and N122D mutations in Sf9 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9189)
  AUTHORS   Mohammadi S, Yang L, Harpak A, Herrera-Alvarez S, Del Pilar 
            Rodriguez-Ordonez M, Peng J, Zhang K, Storz JF, Dobler S, Crawford 
            AJ, Andolfatto P
  TITLE     Concerted evolution reveals co-adapted amino acid substitutions in 
            Na(+)K(+)-ATPase of frogs that prey on toxic toads.
  JOURNAL   Curr Biol. 2021 Jun 21;31(12):2530-2538.e10. doi: 
            10.1016/j.cub.2021.03.089. Epub 2021 Apr 21.
  PUBMED    33887183
REFERENCE   2  (bases 1 to 9189)
  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"; doi: 
            "10.1016/j.cub.2021.03.089"; journalName: "Curr Biol"; date: 
            "2021-06-21- 21"; volume: "31"; issue: "12"; pages: "2530-2538.e10"
COMMENT     Sequence Label: S+Q111R+N122D
FEATURES             Location/Qualifiers
     source          1..9189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    3419..3553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3456..3475)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3510..3529
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3582..3747
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3931..4385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4018..4037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4227..4248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4411..4515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4516..4584,4585..5376)
                     /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     complement(4734..4753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5547..6135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6036..6055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6289..6306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6440..6664
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6731..7264)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7453..7458,7459..7475,7476..7481))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    7989..8037
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     8062..8081
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(9057..9166)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcccggtcc gaagcgcgcg gaattcaaag gcctacgtcg acgagctcac
      181 tagtatggga tacggggccg gacgtgataa gtatgagccc gcagccactt ctgaacatgg
      241 cggcaagaag aagggcaaag ggaaggatag ggatatggat gagctcaaga aggaagtggc
      301 actggatgac cataagatga ccctagatga gctgcatcgc aagtatggaa ccgacctgac
      361 cagggggctt accacagctc gagcagccga aatcctggca cgagatggtc caaatgcctt
      421 gaccccacct ccaaccaccc cagaatgggt gaagttctgc cgccagctgt ttggaggctt
      481 ctctatgctg ctgtggatcg gggccattct ctgcttcctg gcatatggta ttcgggctgc
      541 aacagaggag gagccacaga acgatgatct ctacctcggt gttgtcctgt ccgctgttgt
      601 catcatcact ggctgctttt cctactacca agaagcaaag agctcaaaga tcatggagtc
      661 ctttaagaac atggtgcctc agcaagccct ggttatccga aatggtgaga agcttagtat
      721 caatgcggaa gaggttgtcc agggagatct tgtagaagtg aagggtggtg accggatccc
      781 agccgatctc cgaatcatct ctgctcatgg ctgtaaggtg gacaactctt ccctgactgg
      841 agagtctgag ccccagacca gatctccgga cttcaccaat gagaaccctc tggagacccg
      901 taacatcgcc ttcttctcca ccaactgtgt agaaggaacg gctcgtggta ttgttatcaa
      961 cactggtgac cgcactgtta tgggacgtat tgcaaccttg gcttctggtc ttgagggtgg
     1021 ccgaaccccc attgccattg agattgagca cttcatccat atcatcactg gggtggcagt
     1081 attcctgggt gtctctttct tcatcctgtc actcatcttg cagtacactt ggcttgaagc
     1141 agtcatattc ctcatcggta tcattgtcgc caacgtgcct gaaggtctcc tggctacagt
     1201 cactgtgtgc cttacactca ccgccaagag gatggctcgt aaaaactgct tggtgaagaa
     1261 cttggaagct gtggaaaccc ttggatctac ctcgaccatc tgctctgaca agaccggaac
     1321 cctgactcag aaccgtatga ccgtagcaca catgtggttt gacaaccaga ttcatgaagc
     1381 cgacaccaca gagaatcaga gcggtgcctc ctttgataag agctccccca cctggacagc
     1441 tctggctcgt attgctggtc tctgtaaccg tgcggtgttc caggctggtc aggagaacac
     1501 tcctattctg aagagagatg tggccggaga tgcctctgaa tctgctctgc tgaagtgtat
     1561 tgagctgtgc tgtggctctg taagggacat gcgagaaaag aaccaaaaag tggctgagat
     1621 tcccttcaac tccaccaata aataccagtt gtccgtacac aagaatgcca actcttcaga
     1681 gtctcgatac cttctggtta tgaagggtgc ccccgaaagg atcctggacc gttgctctac
     1741 catcctgctt caaggcaaag agcagcccct agatgaggag ctaaaggacg ctttccagaa
     1801 tgcttacttg gagctcgggg gccttgggga gcgagtactg ggtttctgcc acttagttct
     1861 aaatgacgac cagttccctg aaggcttcaa ctttgataca gaagaagtga atttccccac
     1921 agagggcctg tgctttgttg gccttatctc catgattgac cctccccgcg ccgctgtgcc
     1981 tgatgcagtc ggcaaatgca gaagtgccgg aatcaaggtt atcatggtca ccggtgacca
     2041 ccccatcact gccaaggcta ttgccaaggg agtaggtatc atctctgagg gtaacgagac
     2101 agtggaagac attgcagccc gtctcaacat cccagtcaac caggtgaacc ccagggatgc
     2161 aaaagcatgt gtgatccatg gcacagacct gaaggacatg tcctcggatc agattgacga
     2221 catcctccga caccacacag aaattgtgtt tgccagaacc tcccctcagc agaagctcat
     2281 cattgtggag ggttgtcaga gacagggagc cattgtggca gtgactggtg atggtgtgaa
     2341 cgactccccc gctctgaaga aggcagacat tgggattgct atgggtattg caggctctga
     2401 tgtctccaaa caggctgcag atatgattct gctggacgac aactttgcct ctattgtgac
     2461 tggtgtggag gaagggcgtc tgatttttga taacctgaag aaatctattg cctacacact
     2521 gaccagtaac atccctgaga tcaccccttt cctcatcttt atcattgcca acattcccct
     2581 gcccctgggc acagtcacca tcctctgtat tgatctgggc actgacatgg tccctgccat
     2641 ctccctggcc tatgaacaag cagaaagtga catcatgaag cgtcagccca gaaacccaaa
     2701 gacggacaag ctggtgaatg aacgtctcat cagtatggct tacgggcaga tcggtatgat
     2761 ccaggccctt ggtgggttct tcacctactt tgttatcttg gcggagaacg gtttcctgcc
     2821 atcaacgctg ctcggcatca gggtgaactg ggatgaccga tggataaatg atgtggaaga
     2881 cagctacggg cagcagtgga catacgagca gaggaagatt gtggagttca cctgtcacac
     2941 ctccttcttc gtcagcattg tcattgtaca gtgggctgac ctgatcatct gtaagaccag
     3001 gaggaactct gtattccagc agggcatgaa gaacaagatc cttatctttg gcctgtttga
     3061 ggagactgca ctggccgcct tcctgtcata ctgcccaggc atggatgtgg cactccgcat
     3121 gtacccactg aaaccaacat ggtggttctg tgccttcccg tactcgctgc tcatcttcat
     3181 atatgatgaa atccgaaaac tgataatcag acgcagccct ggcggctggg tggagaaaga
     3241 aacctattat taagcggccg ctttcgaatc tagagcctgc agtctcgaca agcttgtcga
     3301 gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3361 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3421 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3481 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3541 tcatgtctgg atctgatcac tgcttgagcc taggagatcc gaaccagata agtgaaatct
     3601 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     3661 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     3721 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat
     3781 caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac
     3841 agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa
     3901 cgcttatttg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3961 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4021 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4081 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4141 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4201 tgacgttgga gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa
     4261 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4321 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     4381 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4501 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4561 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4621 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4681 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4741 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4801 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4861 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4921 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4981 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5041 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5101 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5161 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5221 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5281 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5341 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6241 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6301 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     6361 tcacaccgca tagaccagcc gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg
     6421 gatgccctgc gtaagcgggt gtgggcggac aataaagtct taaactgaac aaaatagatc
     6481 taaactatga caataaagtc ttaaactaga cagaatagtt gtaaactgaa atcagtccag
     6541 ttatgctgtg aaaaagcata ctggactttt gttatggcta aagcaaactc ttcattttct
     6601 gaagtgcaaa ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg taaagactat
     6661 attcgcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt
     6721 gaacgaattg ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta
     6781 tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga
     6841 tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa
     6901 tgccctgcct ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg
     6961 gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga
     7021 aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg
     7081 gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag
     7141 cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg
     7201 agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta
     7261 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
     7321 gatgcccgag gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact
     7381 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
     7441 tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc
     7501 cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg
     7561 tcctggctgg cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc
     7621 ttgctgttct tctacggcaa ggtgctgtgc acggatctgc ccttgcttca ggagatcggt
     7681 agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc
     7741 ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt
     7801 ggttggctac gtacccgtag tggctatggc agggcttgcc gccccgacgt tggctgcgag
     7861 ccctgggcct tcacccgaac ttgggggttg gggtggggaa aaggaagaaa cgcgggcgta
     7921 ttggtcccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     7981 gcgtttatga acaaacgacc caacacccgt gcgttttatt ctgtcttttt attgccgtca
     8041 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc ccccatctcc
     8101 cggtacctca gctactctta atctccaact ttatatcgaa acgtccctgg aaacggtctt
     8161 tttcgctata ctgtatgttc tggccatatg ccttgcactc gatacgaact tctgtgttta
     8221 agctgatgtt ggtaaattgc acagcaataa ggggttgtag atattgaggt tgtagccgta
     8281 ttccatagta aggatagtag ttcaagggaa atccagcata tccgcccatg ccaaagtagt
     8341 caatcctaaa gatcttaggc tggtcttcct ctttctttcc ttcacagagg attggaatca
     8401 tgttttcact gtacaagcct ttgaaaccaa caactgttcc atttgctgga ggctttggtt
     8461 taaatcccag aattctgtta agcttcacaa tcacacaggg cttgccttct ttaaaaccaa
     8521 aggttggatc atccagacca gagcaatttc cgagccattc cctcttaaat cgacaggctc
     8581 ttttttcgcc tccatctcca tttattggtc cacgttcaat gtaggtagag ggttcagttg
     8641 ggcagtcttc aaatttcaca ttgtcctctt gcttgctgga gttatacttt tctaaaaacc
     8701 tagtaatgct agacacatat tctgcatatg aagccgggtc attctgatta aaggagagtt
     8761 ctgcctttac agctttgggc agctgagtca gtcctggtgg ggcgacacgg tcctgatact
     8821 ttggttcata tatactgata gttaatagaa gcacttgaat ggtcccaata aatataccag
     8881 ctagacatcc atagaatacc agatagaaaa gtaagatttt aaaccagctg ccacctgttc
     8941 tgcccaggaa ctccttcttc tcggggttcc agagaaattt cttccagctg ccgtcgttct
     9001 ccttggtttt gtctctggcc atctcgagat cccgggtgat caagtcttcg tcgagtgatt
     9061 gtaaataaaa tgtaatttac agtatagtat tttaattaat atacaaatga tttgataata
     9121 attcttattt aactataata tattgtgttg ggttgaatta aaggtccgta tactccggaa
     9181 tattaatag
//
