LOCUS       sequence-345188-        9183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the full Leptodactylus latrans Na+K+ATPase with ATP1A1-S 
            variant with 10 R mutations in Sf9 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9183)
  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 9183)
  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+10subs
FEATURES             Location/Qualifiers
     source          1..9183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..153
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     78..95
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    3474..3608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3511..3530)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3565..3584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3637..3802
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3986..4440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4073..4092)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4282..4303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4466..4570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4571..4639,4640..5431)
                     /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(4789..4808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5602..6190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6091..6110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6344..6361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6495..6719
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6786..7319)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7508..7513,7514..7530,7531..7536))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    8044..8092
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     8117..8136
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(join(9112..9183,1..38))
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 ttaactataa tatattgtgt tgggttgaat taaaggtccg tatactccgg aatattaata
       61 gatcatggag ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt
      121 cgtaacagtt ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc
      181 atcgggcgcg gatcccggtc cgaagcgcgc ggaattcaaa atgggatacg gggccggacg
      241 tgataagtat gagcccgcag ccacttctga acatggcggc aagaagaaag gcaaagggaa
      301 ggatagggat atggatgagc tcaagaagga agtggcactg gatgaccata agatgaccct
      361 agatgagctg catcgcaagt atggaaccga cctgaccagg gggcttacca cagctcgagc
      421 agccgaaatc ctggcacgag atggtccaaa tgccttgacc ccacctccaa ccaccccaga
      481 atgggtgaag ttctgccgcc agctgtttgg aggcttctct atgctgctgt ggatcggggc
      541 cattctctgc ttcctggcat atggtattca gactgcaaca gaggatgagc cacagaacga
      601 taatctctac ctcggtgttg tcctgtccgc tgttgtcatc gtcactggct gcttttccta
      661 ctaccaagaa gcaaagagct caaagatcat ggagtccttt aagaacatgg tgcctcagca
      721 agccctggtt atccgaaatg gtgagaagct tagtatcaat gcggaagagg ttgtcctggg
      781 agatcttgta gaagtgaagg gtggtgaccg gatcccagcc gatctccgaa tcctctctgc
      841 tcatggctgt aaggtggaca actcttccct gactggagag tctgagcccc agaccagatc
      901 tccggacttc accaatgaga accctctgga gacccgtaac atcgccttct tctccaccaa
      961 ctgtgtagaa ggaacggctc gtggtattgt tatcaacact ggtgaccgca ctgttatggg
     1021 acgtattgca accttggctt ctggtcttga gggtggccga acccccattg ccgttgagat
     1081 tgagcacttc atccatatca tcactggggt ggcagtattc ctgggtgtct ctttcttcat
     1141 cctgtcactc atcttgcagt acacttggct tgaagcagtc atattcctca tcggtatcat
     1201 tgtcgccaac gtgcctgaag gtctcctggc tacagtcact gtgtgcctta cactcaccgc
     1261 caagaggatg gctcgtaaaa actgcttggt gaagaacttg gaagctgtgg aaacccttgg
     1321 atctacctcg accatctgct ctgacaagac cggaaccctg actcagaacc gtatgaccgt
     1381 agcacacatg tggtttgaca accagattca tgaagccgac accacagaga atcagagcgg
     1441 tgcctgcttt gataagagct cccccacctg gacagctctg gctcgtattg ctggtctctg
     1501 taaccgtgcg gtgttccagg ctggtcagga gaacactcct attctgaaga gagatgtggc
     1561 cggagatgcc tctgaatctg ctctgctgaa gtgtattgag ctgtgctgtg gctctgtaag
     1621 ggacatgcga gaaaagaacc aaaaagtggc tgagattccc ttcaactcca ccaataaata
     1681 ccagttgtcc gtacacaaga atgccaactc ttcagagtct cgataccttc tggttatgaa
     1741 gggtgccccc gaaaggatcc tggaccgttg ctctaccatc ctgcttcaag gcaaagagca
     1801 gcccctagat gaggagctaa aggacgcttt ccagaatgct tacatggagc tcgggggcct
     1861 tggggagcga gtactgggtt tctgccactt agttctaaat gacgaccagt tccctgaagg
     1921 cttcaacttt gatacagaag aagtgaattt ccccacagag ggcctgtgct ttgttggcct
     1981 tatctccatg attgaccctc cccgcgccgc tgtgcctgat gcagtcggca aatgcagaag
     2041 tgccggaatc aaggttatca tggtcaccgg tgaccacccc atcactgcca aggctattgc
     2101 caagggagta ggtatcatct ctgagggtaa cgagacagtg gaagacattg cagcccgtct
     2161 caacatccca gtcaaccagg tgaaccccag ggatgcaaaa gcatgtgtga tccatggcac
     2221 agacctgaag gacatgtcct cggatcagat tgacgacatc ctccgacacc acacagaaat
     2281 tgtgtttgcc agaacctccc ctcagcagaa gctcatcatt gtggagggtt gtcagagact
     2341 gggagccatt gtggcagtga ctggtgatgg tgtgaacgac tcccccgctc tgaagaaggc
     2401 agacattggg attgctatgg gtattgcagg ctctgatgtc tccaaacagg ctgcagatat
     2461 gattctgctg gacgacaact ttgcctctat tgtgactggt gtggaggaag ggcgtctgat
     2521 ttttgataac ctgaagaaat ctattgccta cacactgacc agtaacatcc ctgagatcac
     2581 ccctttcctc atctttatca tggccaacat tcccctgccc ctgggcacag tcaccatcct
     2641 ctgtattgat ctgggcactg acatggtccc tgccatctcc ctggcctatg aacaagcaga
     2701 aagtgacatc atgaagcgtc agcccagaaa cccaaagacg gacaagctgg tgaatgaacg
     2761 tctcatcagt atggcttacg ggcagatcgg tatgatccag gcccttggtg ggttcttcac
     2821 ctactttgtt atcttggcgg agaacggttt cctgccatca acgctgctcg gcatcagggt
     2881 gaactgggat gaccgatgga taaatgatgt ggaagacagc tacgggcagc agtggacata
     2941 cgagcagagg aagattgtgg agttcacctg tcacacctcc ttcttcgtca gcattgtcat
     3001 tgtacagtgg gctgacctga tcatctgtaa gaccaggagg aactctgtat tccagcaggg
     3061 catgaagaac aagatcctta tctttggcct gtttgaggag actgcactgg ccgccttcct
     3121 gtcatactgc ccaggcatgg atgtggcact ccgcatgtac ccactgaaac caacatggtg
     3181 gttctgtgcc ttcccctact ctctgctcat cttcatatat gatgaaatcc gaaaactgat
     3241 aatcagacgc agccctggcg gctgggtgga gaaagaaacc tattattaag tcgacgagct
     3301 cactagtcgc ggccgctttc gaatctagag cctgcagtct cgacaagctt gtcgagaagt
     3361 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3421 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3481 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3541 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3601 tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga aatctagttc
     3661 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
     3721 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
     3781 ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac
     3841 atcctgccaa ctccatgtga caaaccgtca tcttcggcta ctttttctct gtcacagaat
     3901 gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat atcaacgctt
     3961 atttgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4021 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4081 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4141 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4201 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4261 ttggagtcca cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4321 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4381 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4801 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5101 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6361 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     6421 ccgcatagac cagccgcgta acctggcaaa atcggttacg gttgagtaat aaatggatgc
     6481 cctgcgtaag cgggtgtggg cggacaataa agtcttaaac tgaacaaaat agatctaaac
     6541 tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag tccagttatg
     6601 ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat tttctgaagt
     6661 gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg catggtaaag actatattcg
     6721 cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg
     6781 aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc
     6841 aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca
     6901 taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc
     6961 tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc
     7021 tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca
     7081 gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga
     7141 tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc
     7201 atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca
     7261 cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc
     7321 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
     7381 ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc
     7441 gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt
     7501 gcattacagt ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct tcatccgttt
     7561 ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg
     7621 gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt caggcattgg cggccttgct
     7681 gttcttctac ggcaaggtgc tgtgcacgga tctgcccttg cttcaggaga tcggtagacc
     7741 tcggccgtcg cggcgcttgc cggtggtgct gaccccggat gaagtggttc gcatcctcgg
     7801 ttttctggaa ggcgagcatc gtttgttcgc ccaggactct agctatagtt ctagtggttg
     7861 gctacgtacc cgtagtggct atggcagggc ttgccgcccc gacgttggct gcgagccctg
     7921 ggccttcacc cgaacttggg ggttggggtg gggaaaagga agaaacgcgg gcgtattggt
     7981 cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt
     8041 tatgaacaaa cgacccaaca cccgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     8101 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccca tctcccggta
     8161 cctcagctac tcttaatctc caactttata tcgaaacgtc cctggaaacg gtctttttcg
     8221 ctatactgta tgttctggcc atatgccttg cactcgatac gaacttctgt gtttaagctg
     8281 atgttggtaa attgcacagc aataaggggt tgtagatatt gaggttgtag ccgtattcca
     8341 tagtaaggat agtagttcaa gggaaatcca gcatatccgc ccatgccaaa gtagtcaatc
     8401 ctaaagatct taggctggtc ttcctctttc tttccttcac agaggattgg aatcatgttt
     8461 tcactgtaca agcctttgaa accaacaact gttccatttg ctggaggctt tggtttaaat
     8521 cccagaattc tgttaagctt cacaatcaca cagggcttgc cttctttaaa accaaaggtt
     8581 ggatcatcca gaccagagca atttccgagc cattccctct taaatcgaca ggctcttttt
     8641 tcgcctccat ctccatttat tggtccacgt tcaatgtagg tagagggttc agttgggcag
     8701 tcttcaaatt tcacattgtc ctcttgcttg ctggagttat acttttctaa aaacctagta
     8761 atgctagaca catattctgc atatgaagcc gggtcattct gattaaagga gagttctgcc
     8821 tttacagctt tgggcagctg agtcagtcct ggtggggcga cacggtcctg atactttggt
     8881 tcatatatac tgatagttaa tagaagcact tgaatggtcc caataaatat accagctaga
     8941 catccataga ataccagata gaaaagtaag attttaaacc agctgccacc tgttctgccc
     9001 aggaactcct tcttctcggg gttccagaga aatttcttcc agctgccgtc gttctccttg
     9061 gttttgtctc tggccatctc gagatcccgg gtgatcaagt cttcgtcgag tgattgtaaa
     9121 taaaatgtaa tttacagtat agtattttaa ttaatataca aatgatttga taataattct
     9181 tat
//
