LOCUS       sequence-379801-        9287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Na,K-ATPase (ATP1A1 and ATP1B1 in pFastBac Dual vector) in
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9287)
  AUTHORS   Mohammadi S, Ozdemir HI, Ozbek P, Sumbul F, Stiller J, Deng Y, 
            Crawford AJ, Rowland HM, Storz JF, Andolfatto P, Dobler S
  TITLE     Epistatic effects between amino acid insertions and substitutions 
            mediate toxin-resistance of vertebrate Na+, K+-ATPases.
  JOURNAL   Mol Biol Evol. 2022 Dec 6:msac258. doi: 10.1093/molbev/msac258.
  PUBMED    36472530
REFERENCE   2  (bases 1 to 9287)
  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: "Mol Biol 
            Evol. 2022 Dec 6:msac258. doi: 10.1093/molbev/msac258."
COMMENT     Sequence Label: Sandgrouse+R111Q–insD
FEATURES             Location/Qualifiers
     source          1..9287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRLGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4160..4208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4233..4252
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(5237..5346)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5370..5461
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5386..5403
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    8877..9011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8914..8933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8968..8987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9040..9205
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgcccagcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4261 cccccatctc ccggtaccgc atgctcagct gctttttatg tcaaatttaa tatcaaagcg
     4321 tccctggaag cggtctttgt cgctgtactg gatgttctgc ccgtaggccc tgcattccac
     4381 gcgcacctcc atgtcgtagg tcaggttggt gaactgcact gccaccagcg gctgcaggta
     4441 ccgcgactgc aggagcttgc cgtagtaggg atagtactgc agggcaaagc cagggtagcc
     4501 acccatccca tagtactcca ccgtgcctat tttgtctgca tcgtcctctt tcttggcagc
     4561 acagtggaca gggatgagga atgggttgta ttttgccata acctctggag ggaggctgtc
     4621 atttaccgga gccttaggtt tgaagccgat aattctgttg agcttgacga ggatgcatgg
     4681 tttgccatcc ctgtacccaa aggtgggatc atttattcca gaacagtttt ccagccattc
     4741 acgtttgaat ttgcagacct tcttctggcc ctgggcatca ttatatggtc ctctctcttt
     4801 gtaatcgcta ggtatgtccc cacagtcctg aaatatcatg ttttcagtct gttgatcagc
     4861 gctgtagtcc ttcaagaact tctcaaggtt cttcacatat ggctcaaaac ttttggcgtc
     4921 ggagacacta aaggaaattt ctgtcttttg tacctggggt acttgagtca gtcctggagg
     4981 tgccactcga tcctgatatt tgggttcaaa ttcactgaca gtgagcaaca tcacttggat
     5041 ggtcccaatg aatatacctg ccaggcaacc atagaagatc acatagaaga ggaggatttt
     5101 aaaccagctg cccccggtgc ggcccagcag ctccttcttc tccgagttcc agatgaactt
     5161 cttccagttg ccgtcgccgt ccttggcctt cccgcgggcc atctcgagat cccgggtgat
     5221 caagtcttcg tcgagtgatt gtaaataaaa tgtaatttac agtatagtat tttaattaat
     5281 atacaaatga tttgataata attcttattt aactataata tattgtgttg ggttgaatta
     5341 aaggtccgta tactccggaa tattaataga tcatggagat aattaaaatg ataaccatct
     5401 cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa aacctataaa
     5461 tattccggat tattcatacc gtcccaccat cgggcgcgga tcccggtccg aagcgcgcgg
     5521 aattcaaagg cctacgtcga cgagctcacc tatagggcga attggcggaa ggccgtcaag
     5581 gccgcatgtc gacatgggca agggggctgg aagagacaag tatgagccta ctgcaacatc
     5641 cgaacatggc acaaagaaga aaaaggggaa gaaggagagg gacatggatg aacttaaaaa
     5701 ggaagtcaca atggatgacc acaagctgag ccttgatgaa cttcatcgta aatatggaac
     5761 agacttgagt aggggtttga ctactgcacg tgcagctgag attttggctc gtgatggccc
     5821 aaattccctc acacccccac ccaccactcc tgaatggctc aagttctgcc ggcagctctt
     5881 tggaggattc tcactcctgt tgtggattgg tgctgttctg tgctttctgg cttatggcat
     5941 acaaagtgcg gtgcaggagg cggccgacaa tgataatctg tacttgggta ttgtgttggc
     6001 agctgtggtt atcattactg gctgtttctc ttattaccaa gaagcaaaaa gttccaagat
     6061 catggagtcc ttcaagaaca tggtgcctca gcaagctctt gtagtcagaa atggtgagaa
     6121 gatgagcata aatgctgaag gtgttgtagt tggagatcta gtggaggtga aaggaggaga
     6181 cagaattcca gccgaccttc ggatcatatc tgcacatggt tgcaaggtag ataactcctc
     6241 acttactggt gaatcagagc ctcagaccag atctccagac ttctccaatg agaatcccct
     6301 ggagaccagg aacattgcct tcttttccac caactgtgtg gaaggcactg cccgtggcat
     6361 tgtcattagc actggggatc gcactgtaat gggccgtatt gccagtctgg cttctgggct
     6421 ggaagggggg aaaactccaa ttgccatgga gattgagcac tttatccacc tcatcactgg
     6481 agtagctgtc ttcctgggtg tgactttctt catcctttca ctcatccttg agtacacatg
     6541 gctggaggct gtaatcttcc tcatcgggat cattgttgcc aatgtccctg aagggctgct
     6601 tgcaacagtc acggtatgtc tgacactaac agccaagcgt atggctcgta agaactgctt
     6661 ggtgaagaac ctcgaagctg tggaaaccct gggttccacg tccactatct gttctgacaa
     6721 aacaggcacg ctgacacaga atcgcatgac ggttgcgcac atgtggtttg acaatcagat
     6781 tcatgaggct gatactacag agaaccagag tggtgcttcc tttgacaaga gctcagctac
     6841 ttggactgct ttgtccagag ttgcaggtct ctgcaaccgt gctgtgtttc aggccaacca
     6901 ggaaaatgta ccaatactta agagagcagt ggcgggagat gcctctgagt ctgcacttct
     6961 gaaatgcatt gaattgtgct gtggtcctgt caaggagatg agagaaaggt atcccaaagt
     7021 ggtggaaata ccatttaact ctaccaacaa gtatcagctg tctatccaca aaaatgcaaa
     7081 tccgtcagaa tcccgttact tactggtgat gaagggagct ccagagagga ttttggatcg
     7141 ctgcagcacc attcttattc atggcaaaga gcaaccactg gatgaggaaa tgaaagattc
     7201 ttttcaaaat gcctaccttg agttgggagg ccttggggag agagtgttag gattctgtca
     7261 cttggctctg cctgatgatc agttccctga tggcttccag tttgatacag acgaagtgaa
     7321 ctttcctgta gaaaaactct gctttgtagg actgatgtct atgattgacc ctcctcgtgc
     7381 tgctgtgcca gatgctgttg gcaaatgcag aagcgctggg atcaaggtta tcatggttac
     7441 gggagaccat ccgatcacag ccaaagccat tgccaagggt gtcggcatca tctctgaggg
     7501 caatgaaaca gttgaagata ttgctgctcg actcaacatt cctgtcagcc aggtcaaccc
     7561 tagggatgcc aaagcttgtg ttgttcatgg ctcagatttg aaggacatga ctagtgagca
     7621 actggatgac atcttgcttc atcatactga aattgtcttt gccaggacat ctcctcagca
     7681 gaagcttata attgtggaag gctgtcagcg acagggtgcc attgtagcag tcacaggtga
     7741 tggtgtgaat gattccccag ccctgaagaa ggctgacatt ggtgttgcta tgggtattgc
     7801 tggctcagat gtctccaagc aggcagctga catgattctg ctggatgata actttgcctc
     7861 cattgtcact ggggttgaag aagggcgtct gatctttgat aacctgaaga agtctattgc
     7921 ttacaccttg accagtaaca ttcctgaaat cacgccgttc ctgatcttca tcattgcaaa
     7981 catacccctt ccactgggaa cagtcaccat cctctgcatt gacttgggca ctgacatggt
     8041 ccctgctatc tccctggcat atgagcaagc agagagtgac atcatgaaga ggcagcccag
     8101 aaatcccaaa acagacaagc tggttaatga acggctgatc agcatggcct atgggcagat
     8161 tggtatgatc caggcccttg gaggtttctt cacctatttt gtaatcatgg cagagaatgg
     8221 gttctggcct tctggcttgc tagggatcag agttcagtgg gatgaccgat gggttaacga
     8281 tgtggaagac agctatgggc agcagtggac ctacgaacag aggaaaatag tggagttcac
     8341 ttgccataca gccttctttg tcagcattgt ggttgtgcag tgggcagact tgatcatttg
     8401 taagaccaga agaaactctg tcttccagca gggaatgaag aacaagatct taatatttgg
     8461 gctctttgag gagactgctc tggctgcctt cctctcctac tgccctggaa tggatgttgc
     8521 tctaaggatg taccctctca agccaacctg gtggttctgc gctttcccat actctcttct
     8581 catattcctg tatgatgaaa tcagaaagct cattatcaga cgcaaccctg gtggttgggt
     8641 ggaaaaagag acctactact aagcggccgc ctgggcctca tgggccttcc gctcactgcc
     8701 cgctttccag tgcggccgct ttcgaatcta gagcctgcag tctcgacaag cttgtcgaga
     8761 agtactagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     8821 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     8881 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     8941 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     9001 atgtctggat ctgatcactg cttgagccta ggagatccga accagataag tgaaatctag
     9061 ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc
     9121 catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag
     9181 ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca
     9241 aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctacttt
//
