LOCUS       sequence-334483-        9195 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the full Leptodactylus latrans Na+K+ATPase with ATP1A1-R 
            variant with reverted Q111R and N122D mutations in Sf9 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9195)
  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 9195)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: R-Q111R-N122D
FEATURES             Location/Qualifiers
     source          1..9195
                     /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    3425..3559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3462..3481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3516..3535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3588..3753
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3937..4391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4024..4043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4233..4254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4417..4521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4522..4590,4591..5382)
                     /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(4740..4759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5553..6141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6042..6061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6295..6312
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6446..6670
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6737..7270)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7459..7464,7465..7481,7482..7487))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    7995..8043
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     8068..8087
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(9063..9172)
                     /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 gacgtgatga gtatgagccc gcagccactt ctgaacatgg
      241 cggcaagaag aaaggcaaag ggaaggataa ggataaggat atggatgagc tcaagaagga
      301 agtggcactg gatgaccata agatgaccct agatgagctg catcgcaagt atggaaccga
      361 cctgaccagg gggcttacca cagaacgggc agccgaaatc ctggcacgag atggtccaaa
      421 tgccttgacc ccacctccaa ccaccccaga atgggtgaag ttctgccgcc agctgtttgg
      481 aggcttctct atgctgctgt ggatcggggc ctttctctgc ttcctggcat atggtattca
      541 gactgcaact gaggatgagc cacaaaatga taatctctac ctcggtgttg tactgtccgc
      601 tgtcgtcata gtcactggct gcttttccta ctaccaagaa gcaaagagct caaagatcat
      661 ggagtccttt aagaacatgg tgcctcagca agccctggtt atccgaaatg gtgagaagct
      721 tagtatcaat gcagaagagg ttgtcctggg agatcttgta gaagtgaagg gtggtgaccg
      781 gatcccagcc gatctccgta tactctctgc ccatggctgt aaggtggaca actcttccct
      841 gactggagag tctgagcccc agaccagatc tccggacttc accaatgaga accctctgga
      901 gacccgtaac atcgccttct tctccaccaa ctgtgtagaa ggaacggctc gtggtattgt
      961 tatcaacact ggtgaccgca ctgttatggg acgtattgca accttggctt ctggtcttga
     1021 gggtggccga acccccattg ctgttgagat tgagcacttc atccatatca tcactggggt
     1081 ggcagtcttc ctgggtgtct ctttcttcgt cctgtcactc atcttgcagt acacttggct
     1141 tgaagcagtc atattcctca tcggtatcat tgtcgccaac gtgcctgaag gtctcctggc
     1201 tacagtcact gtgtgcctta cactcaccgc caagaggatg gctcgtaaaa actgcttggt
     1261 gaagaacttg gaagcagtgg aaacccttgg atctacctcg accatctgct ctgacaagac
     1321 cggaaccctg actcagaacc gtatgaccgt agcacacatg tggtttgaca accagattca
     1381 tgaagccgac accacagaga atcagagcgg tgcctgcttt gataagagct cccccacctg
     1441 gacagctctg gctcgtattg ctggtctttg taaccgtgcg gtgttccagg ctggtcaaga
     1501 gaacactcct attctgaaga gagatgtggc cggagatgcc tcagaatctg ctctgctgaa
     1561 gtgtattgag ctgtgctgtg gctcggtaag ggacatgcga gaaaagaacc aaaaagtggc
     1621 tgagattccc ttcaactcca ccaataaata ccagttgtcc atacacaaga atgccaactc
     1681 ttcagagtct cgataccttc tggttatgaa gggtgccccc gaaaggatcc tggaccgttg
     1741 ctctaccatc ctgcttcaag gcaaagagca gcccctagat gaggagctaa aggacgcttt
     1801 ccagaatgct tacatggagc tcgggggcct tggggagcga gtactgggtt tctgccactt
     1861 agttctaaat gacgaccagt tccctgaagg cttcaacttt gatacagaag aagtgaattt
     1921 ccccacagag ggcctgtgct ttgttggcct tatctccatg attgaccctc cccgcgccgc
     1981 tgtgcctgat gcagtcggca aatgcagaag tgccggaatc aaggttatca tggtcaccgg
     2041 tgaccacccc atcactgcca aggctattgc caagggagta ggtatcatct ctgagggtaa
     2101 cgagacagtg gaagacattg cagcccgtct caacatccca gtcaaccagg tgaaccccag
     2161 ggatgcaaaa gcatgtgtga tccatggcac agacctgaag gacatgtcct tggaacagat
     2221 tgacgacatc ctccgacacc acacagaaat tgtgtttgcc agaacctccc ctcagcagaa
     2281 gctcatcatt gtggagggtt gtcagcgact gggagccatt gtggcagtga ctggtgatgg
     2341 tgtgaacgac tcccccgctc tgaagaaggc agacattggg attgctatgg gtattgcagg
     2401 ctctgatgtc tccaaacagg ctgcagatat gattctgctg gacgacaact ttgcctctat
     2461 tgtgactggt gtggaggaag ggcgtctgat ttttgataac ctgaagaaat ctattgccta
     2521 cacactgacc agtaacatcc ctgagatcac ccctttcctc atctttatca tggccaacat
     2581 tcccctgccc ctgggcacag tcaccatcct ctgtattgat ctgggcactg acatggtccc
     2641 tgccatctcc ctggcctatg aacaagcaga aagtgacatc atgaagcgtc agcccagaaa
     2701 cccaaagacg gacaagctgg tgaatgagcg tctcatcagt atggcttacg ggcagatcgg
     2761 tatgatccag gcccttggtg ggttcttcac ctactttgtt atcttggcgg agaacggttt
     2821 cctgccatca acgctgctcg gcatcagggt aaactgggat gaccgatggg taaatgatgt
     2881 ggaagacagc tacgggcagc agtggaccta cgagcagagg aagattgtgg agttcacctg
     2941 tcacacctcc ttcttcgtca gcattgtcat tgtacagtgg gctgacctga tcatctgtaa
     3001 gaccaggagg aactctgtat tccagcaggg catgaagaac aagatcctta tctttggcct
     3061 gtttgaggag actgcactgg ccgccttcct gtcatactgc ccaggcatgg atgtggcact
     3121 ccgcatgtac ccactgaaac caacatggtg gttctgtgcc ttcccctact ctctgctcat
     3181 cttcatatat gatgaaatcc gaaaactgat aatcagacgc agccctggcg gctgggtgga
     3241 gaaagaaacc tattattaag cggccgcttt cgaatctaga gcctgcagtc tcgacaagct
     3301 tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3361 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3421 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3481 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3541 atcttatcat gtctggatct gatcactgct tgagcctagg agatccgaac cagataagtg
     3601 aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac
     3661 ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc
     3721 cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt
     3781 tttaatcaaa catcctgcca actccatgtg acaaaccgtc atcttcggct actttttctc
     3841 tgtcacagaa tgaaaatttt tctgtcatct cttcgttatt aatgtttgta attgactgaa
     3901 tatcaacgct tatttgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3961 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4021 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4081 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4141 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4201 gccctttgac gttggagtcc acgttcttaa tagtggactc ttgttccaaa ctggaacaac
     4261 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4381 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4441 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4501 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4561 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4621 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4681 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4741 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4801 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4861 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4921 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4981 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5041 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5101 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5161 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5221 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5281 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5341 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6181 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6241 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6301 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     6361 ggtatttcac accgcataga ccagccgcgt aacctggcaa aatcggttac ggttgagtaa
     6421 taaatggatg ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa ctgaacaaaa
     6481 tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca
     6541 gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca
     6601 ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg gcatggtaaa
     6661 gactatattc gcggcgttgt gacaatttac cgaacaactc cgcggccggg aagccgatct
     6721 cggcttgaac gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt
     6781 cccgtatgcc caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca
     6841 cgtagatcac ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg
     6901 tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag atcatagata tagatctcac
     6961 tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt
     7021 ggtcgaaggc agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg
     7081 agtccggctg atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa
     7141 gagcagcccg catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca
     7201 tacttgagcc acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc
     7261 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     7321 tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc
     7381 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc
     7441 atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc
     7501 ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca
     7561 tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg
     7621 gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctt gcttcaggag
     7681 atcggtagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga tgaagtggtt
     7741 cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc tagctatagt
     7801 tctagtggtt ggctacgtac ccgtagtggc tatggcaggg cttgccgccc cgacgttggc
     7861 tgcgagccct gggccttcac ccgaacttgg gggttggggt ggggaaaagg aagaaacgcg
     7921 ggcgtattgg tcccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg
     7981 aaccccgcgt ttatgaacaa acgacccaac acccgtgcgt tttattctgt ctttttattg
     8041 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     8101 atctcccggt acctcagcta ctcttaatct ccaactttat atcgaaacgt ccctggaaac
     8161 ggtctttttc gctatactgt atgttctggc catatgcctt gcactcgata cgaacttctg
     8221 tgtttaagct gatgttggta aattgcacag caataagggg ttgtagatat tgaggttgta
     8281 gccgtattcc atagtaagga tagtagttca agggaaatcc agcatatccg cccatgccaa
     8341 agtagtcaat cctaaagatc ttaggctggt cttcctcttt ctttccttca cagaggattg
     8401 gaatcatgtt ttcactgtac aagcctttga aaccaacaac tgttccattt gctggaggct
     8461 ttggtttaaa tcccagaatt ctgttaagct tcacaatcac acagggcttg ccttctttaa
     8521 aaccaaaggt tggatcatcc agaccagagc aatttccgag ccattccctc ttaaatcgac
     8581 aggctctttt ttcgcctcca tctccattta ttggtccacg ttcaatgtag gtagagggtt
     8641 cagttgggca gtcttcaaat ttcacattgt cctcttgctt gctggagtta tacttttcta
     8701 aaaacctagt aatgctagac acatattctg catatgaagc cgggtcattc tgattaaagg
     8761 agagttctgc ctttacagct ttgggcagct gagtcagtcc tggtggggcg acacggtcct
     8821 gatactttgg ttcatatata ctgatagtta atagaagcac ttgaatggtc ccaataaata
     8881 taccagctag acatccatag aataccagat agaaaagtaa gattttaaac cagctgccac
     8941 ctgttctgcc caggaactcc ttcttctcgg ggttccagag aaatttcttc cagctgccgt
     9001 cgttctcctt ggttttgtct ctggccatct cgagatcccg ggtgatcaag tcttcgtcga
     9061 gtgattgtaa ataaaatgta atttacagta tagtatttta attaatatac aaatgatttg
     9121 ataataattc ttatttaact ataatatatt gtgttgggtt gaattaaagg tccgtatact
     9181 ccggaatatt aatag
//
