LOCUS       sequence-334485-        9183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the full Leptodactylus latrans Na+K+ATPase with ATP1A1-S 
            variant with 12 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+12subs
FEATURES             Location/Qualifiers
     source          1..9183
                     /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    3413..3547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3450..3469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3504..3523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3576..3741
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3925..4379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4012..4031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4221..4242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4405..4509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4510..4578,4579..5370)
                     /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(4728..4747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5541..6129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6030..6049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6283..6300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6434..6658
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6725..7258)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7447..7452,7453..7469,7470..7475))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    7983..8031
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     8056..8075
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(9051..9160)
                     /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 gaattcaaaa tgggatacgg ggccggacgt
      181 gataagtatg agcccgcagc cacttctgaa catggcggca agaagaaagg caaagggaag
      241 gatagggata tggatgagct caagaaggaa gtggcactgg atgaccataa gatgacccta
      301 gatgagctgc atcgcaagta tggaaccgac ctgaccaggg ggcttaccac agctcgagca
      361 gccgaaatcc tggcacgaga tggtccaaat gccttgaccc cacctccaac caccccagaa
      421 tgggtgaagt tctgccgcca gctgtttgga ggcttctcta tgctgctgtg gatcggggcc
      481 attctctgct tcctggcata tggtattcgt actgcaacag aggatgagcc acagaacgat
      541 gatctctacc tcggtgttgt cctgtccgct gttgtcatcg tcactggctg cttttcctac
      601 taccaagaag caaagagctc aaagatcatg gagtccttta agaacatggt gcctcagcaa
      661 gccctggtta tccgaaatgg tgagaagctt agtatcaatg cggaagaggt tgtcctggga
      721 gatcttgtag aagtgaaggg tggtgaccgg atcccagccg atctccgaat cctctctgct
      781 catggctgta aggtggacaa ctcttccctg actggagagt ctgagcccca gaccagatct
      841 ccggacttca ccaatgagaa ccctctggag acccgtaaca tcgccttctt ctccaccaac
      901 tgtgtagaag gaacggctcg tggtattgtt atcaacactg gtgaccgcac tgttatggga
      961 cgtattgcaa ccttggcttc tggtcttgag ggtggccgaa cccccattgc cgttgagatt
     1021 gagcacttca tccatatcat cactggggtg gcagtattcc tgggtgtctc tttcttcatc
     1081 ctgtcactca tcttgcagta cacttggctt gaagcagtca tattcctcat cggtatcatt
     1141 gtcgccaacg tgcctgaagg tctcctggct acagtcactg tgtgccttac actcaccgcc
     1201 aagaggatgg ctcgtaaaaa ctgcttggtg aagaacttgg aagctgtgga aacccttgga
     1261 tctacctcga ccatctgctc tgacaagacc ggaaccctga ctcagaaccg tatgaccgta
     1321 gcacacatgt ggtttgacaa ccagattcat gaagccgaca ccacagagaa tcagagcggt
     1381 gcctgctttg ataagagctc ccccacctgg acagctctgg ctcgtattgc tggtctctgt
     1441 aaccgtgcgg tgttccaggc tggtcaggag aacactccta ttctgaagag agatgtggcc
     1501 ggagatgcct ctgaatctgc tctgctgaag tgtattgagc tgtgctgtgg ctctgtaagg
     1561 gacatgcgag aaaagaacca aaaagtggct gagattccct tcaactccac caataaatac
     1621 cagttgtccg tacacaagaa tgccaactct tcagagtctc gataccttct ggttatgaag
     1681 ggtgcccccg aaaggatcct ggaccgttgc tctaccatcc tgcttcaagg caaagagcag
     1741 cccctagatg aggagctaaa ggacgctttc cagaatgctt acatggagct cgggggcctt
     1801 ggggagcgag tactgggttt ctgccactta gttctaaatg acgaccagtt ccctgaaggc
     1861 ttcaactttg atacagaaga agtgaatttc cccacagagg gcctgtgctt tgttggcctt
     1921 atctccatga ttgaccctcc ccgcgccgct gtgcctgatg cagtcggcaa atgcagaagt
     1981 gccggaatca aggttatcat ggtcaccggt gaccacccca tcactgccaa ggctattgcc
     2041 aagggagtag gtatcatctc tgagggtaac gagacagtgg aagacattgc agcccgtctc
     2101 aacatcccag tcaaccaggt gaaccccagg gatgcaaaag catgtgtgat ccatggcaca
     2161 gacctgaagg acatgtcctc ggatcagatt gacgacatcc tccgacacca cacagaaatt
     2221 gtgtttgcca gaacctcccc tcagcagaag ctcatcattg tggagggttg tcagagactg
     2281 ggagccattg tggcagtgac tggtgatggt gtgaacgact cccccgctct gaagaaggca
     2341 gacattggga ttgctatggg tattgcaggc tctgatgtct ccaaacaggc tgcagatatg
     2401 attctgctgg acgacaactt tgcctctatt gtgactggtg tggaggaagg gcgtctgatt
     2461 tttgataacc tgaagaaatc tattgcctac acactgacca gtaacatccc tgagatcacc
     2521 cctttcctca tctttatcat ggccaacatt cccctgcccc tgggcacagt caccatcctc
     2581 tgtattgatc tgggcactga catggtccct gccatctccc tggcctatga acaagcagaa
     2641 agtgacatca tgaagcgtca gcccagaaac ccaaagacgg acaagctggt gaatgaacgt
     2701 ctcatcagta tggcttacgg gcagatcggt atgatccagg cccttggtgg gttcttcacc
     2761 tactttgtta tcttggcgga gaacggtttc ctgccatcaa cgctgctcgg catcagggtg
     2821 aactgggatg accgatggat aaatgatgtg gaagacagct acgggcagca gtggacatac
     2881 gagcagagga agattgtgga gttcacctgt cacacctcct tcttcgtcag cattgtcatt
     2941 gtacagtggg ctgacctgat catctgtaag accaggagga actctgtatt ccagcagggc
     3001 atgaagaaca agatccttat ctttggcctg tttgaggaga ctgcactggc cgccttcctg
     3061 tcatactgcc caggcatgga tgtggcactc cgcatgtacc cactgaaacc aacatggtgg
     3121 ttctgtgcct tcccctactc tctgctcatc ttcatatatg atgaaatccg aaaactgata
     3181 atcagacgca gccctggcgg ctgggtggag aaagaaacct attattaagt cgacgagctc
     3241 actagtcgcg gccgctttcg aatctagagc ctgcagtctc gacaagcttg tcgagaagta
     3301 ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3361 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3421 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3481 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3541 ctggatctga tcactgcttg agcctaggag atccgaacca gataagtgaa atctagttcc
     3601 aaactatttt gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc
     3661 tattttgtca ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc
     3721 caactatttt gtccgcccac agcggggcat ttttcttcct gttatgtttt taatcaaaca
     3781 tcctgccaac tccatgtgac aaaccgtcat cttcggctac tttttctctg tcacagaatg
     3841 aaaatttttc tgtcatctct tcgttattaa tgtttgtaat tgactgaata tcaacgctta
     3901 tttgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3961 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4021 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4081 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4141 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4201 tggagtccac gttcttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4261 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4321 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     4381 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4441 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4501 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4561 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4621 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4681 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4741 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4801 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4861 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4921 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4981 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5041 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5101 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5161 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5221 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5281 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5341 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5401 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5461 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5521 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5641 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5701 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5761 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5821 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5881 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5941 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6001 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6061 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6121 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6181 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6241 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6301 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     6361 cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatgcc
     6421 ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact gaacaaaata gatctaaact
     6481 atgacaataa agtcttaaac tagacagaat agttgtaaac tgaaatcagt ccagttatgc
     6541 tgtgaaaaag catactggac ttttgttatg gctaaagcaa actcttcatt ttctgaagtg
     6601 caaattgccc gtcgtattaa agaggggcgt ggccaagggc atggtaaaga ctatattcgc
     6661 ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga
     6721 attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca
     6781 actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat
     6841 aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct
     6901 gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct
     6961 caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag
     7021 caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat
     7081 gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca
     7141 tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac
     7201 ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg
     7261 ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc
     7321 cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg
     7381 ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg
     7441 cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcctt catccgtttc
     7501 cacggtgtgc gtcacccggc aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg
     7561 ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc ggccttgctg
     7621 ttcttctacg gcaaggtgct gtgcacggat ctgcccttgc ttcaggagat cggtagacct
     7681 cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg aagtggttcg catcctcggt
     7741 tttctggaag gcgagcatcg tttgttcgcc caggactcta gctatagttc tagtggttgg
     7801 ctacgtaccc gtagtggcta tggcagggct tgccgccccg acgttggctg cgagccctgg
     7861 gccttcaccc gaacttgggg gttggggtgg ggaaaaggaa gaaacgcggg cgtattggtc
     7921 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     7981 atgaacaaac gacccaacac ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     8041 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccat ctcccggtac
     8101 ctcagctact cttaatctcc aactttatat cgaaacgtcc ctggaaacgg tctttttcgc
     8161 tatactgtat gttctggcca tatgccttgc actcgatacg aacttctgtg tttaagctga
     8221 tgttggtaaa ttgcacagca ataaggggtt gtagatattg aggttgtagc cgtattccat
     8281 agtaaggata gtagttcaag ggaaatccag catatccgcc catgccaaag tagtcaatcc
     8341 taaagatctt aggctggtct tcctctttct ttccttcaca gaggattgga atcatgtttt
     8401 cactgtacaa gcctttgaaa ccaacaactg ttccatttgc tggaggcttt ggtttaaatc
     8461 ccagaattct gttaagcttc acaatcacac agggcttgcc ttctttaaaa ccaaaggttg
     8521 gatcatccag accagagcaa tttccgagcc attccctctt aaatcgacag gctctttttt
     8581 cgcctccatc tccatttatt ggtccacgtt caatgtaggt agagggttca gttgggcagt
     8641 cttcaaattt cacattgtcc tcttgcttgc tggagttata cttttctaaa aacctagtaa
     8701 tgctagacac atattctgca tatgaagccg ggtcattctg attaaaggag agttctgcct
     8761 ttacagcttt gggcagctga gtcagtcctg gtggggcgac acggtcctga tactttggtt
     8821 catatatact gatagttaat agaagcactt gaatggtccc aataaatata ccagctagac
     8881 atccatagaa taccagatag aaaagtaaga ttttaaacca gctgccacct gttctgccca
     8941 ggaactcctt cttctcgggg ttccagagaa atttcttcca gctgccgtcg ttctccttgg
     9001 ttttgtctct ggccatctcg agatcccggg tgatcaagtc ttcgtcgagt gattgtaaat
     9061 aaaatgtaat ttacagtata gtattttaat taatatacaa atgatttgat aataattctt
     9121 atttaactat aatatattgt gttgggttga attaaaggtc cgtatactcc ggaatattaa
     9181 tag
//
