LOCUS       sequence-379797-        9293 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 9293)
  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 9293)
  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: Chinchilla+N122D–insM
FEATURES             Location/Qualifiers
     source          1..9293
                     /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
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     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(5240..5349)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5373..5464
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5389..5406
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    8883..9017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8920..8939)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8974..8993
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9046..9211
                     /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 atgccctgcc 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 cttaacttca atttttacat caaagcgtcc
     4321 ctgaaaacgg tctttctcac tgtacccaat gttctcaccg tacgccttac actcaatgcg
     4381 aacttcagtg tccatggtga ggttggtgaa ctgtacggcc agcaggggct gcagatactt
     4441 gggctgcagg agtttgccgt agtaggggta atactgcaga ggaaacccgg cgaatccgcc
     4501 cattccaaag tattccacag ttccaatttt atctttatca tcttctcgct tgccagtgca
     4561 ctgaaccggc agaacatacg ggttgtactt catccctgcg taaatatccg tggactccaa
     4621 agactcattc ttgggaggct ttggtttgaa acccagaact cggttgagct ttatgatgat
     4681 gcagggtttg ccatctctgt agccgtaagt ctcatcattc attccagagc agtttcccag
     4741 ccattcgagc ttgaacctgc agacctttcg ctcaccccgt tcatggctgt agtctcctcg
     4801 ttctttgggt tcactgggca cagtgccaca gtcttcgaaa atcatgtcat ccttctgggc
     4861 tgcatctttg tacttgtcca ggaacctaat tatgttcaac acataggcct catagctctt
     4921 gggatcatta ggacgaaagg aaatttcagt cttctggatc tgaggaatct gtgttaatcc
     4981 tggtggggcc acccggtcct ggtaggtggg cttcagttca ctgatggtca gcaacatcac
     5041 ctggatggtt ccgatgaaga tgccagccag gcagccgtaa aatatcacgt agaacaggag
     5101 gatcttaaac cagctgccgc cggtccggcc caggaactcc ttcttctccg agttccagat
     5161 gaacttcttc cagctgccct cctccttggc tttcccgcgg gccatctcga gatcccgggt
     5221 gatcaagtct tcgtcgagtg attgtaaata aaatgtaatt tacagtatag tattttaatt
     5281 aatatacaaa tgatttgata ataattctta tttaactata atatattgtg ttgggttgaa
     5341 ttaaaggtcc gtatactccg gaatattaat agatcatgga gataattaaa atgataacca
     5401 tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat
     5461 aaatattccg gattattcat accgtcccac catcgggcgc ggatcccggt ccgaagcgcg
     5521 cggaattcaa aggcctacgt cgacgagctc acctataggg cgaattggcg gaaggccgtc
     5581 aaggccgcat gtcgacatgg ggaagggggt tggacgtgat aaatatgaac ccgcagccgt
     5641 ttcagaacaa ggcgacaaaa agagcagaaa ggccaagaaa gagagggata tggatgaact
     5701 gaagaaagaa gtttctatgg atgaccataa acttagcctt gatgaacttc atcgtaaata
     5761 tggaacagac ttgagccgcg gcttaaccgc tgctcgggct gctgagatcc tggcccgaga
     5821 tggtcctaat tccctcacac cccctcccac tacacctgaa tgggtcaaat tctgccggca
     5881 gctgtttgga gggttctcga tgttactgtg gattggagca atactttgct tcttggccta
     5941 tggcattgaa agtttaagcg aagagggacc ttcaaacgat gatctgtatc ttggtgtggt
     6001 gctgtctgct gtcgtcatca taactggctg tttctcctac tatcaagaag ctaaaagttc
     6061 aaagattatg gattccttca aaaacatggt ccctcagcaa gctctagtga ttcgaaatgg
     6121 tgagaaaatg agcatcaatg cagaagatgt tgtagttggg gatctggtgg aagtaaaagg
     6181 aggagacaga atccctgctg atctccgaat catatctgca aatggctgca aggtggataa
     6241 ctcctccctc actggagaat cagaacccca gactaggtct ccggatttca caaatgaaaa
     6301 tcccttggag accaggaata ttgccttttt ttcaaccaac tgtgtcgaag gcactgctcg
     6361 tggcattgtt gtgtacactg gggaccgcac tgtgatggga aggattgcta ccctggcttc
     6421 tgggctagaa ggaggccaga cacccattgc tgcagaaatt gaacatttca tccacatcat
     6481 cacgggtgtg gctgtgttcc tgggagtgtc tttcttcatc ctttctctaa tcctcgagta
     6541 cacctggctt gaggctgtca tcttcctcat tggcattatc gtagccaatg tgccagaagg
     6601 attgctggcc actgtcacgg tgtgtctgac gcttactgcc aaacgcatgg cgcggaagaa
     6661 ctgcttggtg aagaacttag aagctgtgga aacgttggga tccacatcta ccatctgctc
     6721 agataaaact ggaactctga ctcaaaaccg aatgacagtg gcccacatgt ggtttgacaa
     6781 tcaaatccat gaagctgata caacagagaa tcagagcggt gtctcattcg acaagacctc
     6841 agccacctgg cttgctctgt ccagaattgc aggtctttgt aacagggcag tgtttcaggc
     6901 taaccaggaa aacataccta ttcttaagcg ggcagttgca ggagatgcct ctgagtcagc
     6961 actgttaaaa tgcattgagc tgtgctgtgg atctgtgaag gatatgagag acagatatgc
     7021 caagatcgtg gagataccgt tcaactccac caacaagtac cagttatcca ttcataagaa
     7081 ccccaacaca aacgagccac gacacctgtt ggtgatgaag ggcgctccgg agaggatcct
     7141 ggaccggtgc aactctatcc tcctccatgg caaggaacag cccctggacg aggagctgaa
     7201 ggatgctttt cagaacgcct acctggagct gggtggcctt ggagagcgag tgctgggttt
     7261 ctgccacctt catctaccag atgaacagtt tccagaaggg ttccagtttg acactgatga
     7321 agtgaatttc cccctggata atctctgctt tgttgggctc atctccatga tcgacccccc
     7381 tcgggctgct gttcctgatg ccgtgggcaa atgtcgaagc gcgggaatta aggtcatcat
     7441 ggtcacggga gaccatccaa tcacagcgaa agccattgcc aaaggcgtgg gcatcatctc
     7501 agagggcaac gagactgtgg aagacatcgc tgcccgcctc aacatcccag tgagccaggt
     7561 gaaccccaga gatgccaagg cctgtgtcct acatggcagt gacctgaagg acatgagccc
     7621 agagcagctg gatgacattt tgaagcacca cactgagatt gtgtttgcca ggacatcccc
     7681 tcaacagaag cttatcattg tggaaggctg ccagagacag ggtgctattg tggccgtcac
     7741 tggtgatggt gtcaatgact ctccagcttt gaagaaagca gacattgggg ttgctatggg
     7801 gattgcaggc tccgacgtgt ctaaacaagc tgctgacatg attcttttgg atgacaactt
     7861 tgcctcaatt gtgactggag tagaagaagg tcgtctgatc tttgataact tgaagaaatc
     7921 cattgcttac accctaacca gtaacattcc agagatcacc cccttcctga tatttattat
     7981 tgcaaacatt ccactaccac tggggactgt caccatcctc tgcattgact taggcactga
     8041 catggtccct gccatctcac tggcgtatga gcaagctgag agtgacatca tgaaaagaca
     8101 gcccagaaac cccaaaactg acaagctcgt gaacgagcgg ctgatcagca tggcctatgg
     8161 acagatcggt atgatccagg ccctcggggg cttcttcact tactttgtga ttctggctga
     8221 gaatggcttc cttccaattc acttgttggg cctccgtgtg gactgggatg accgctggat
     8281 caatgacatg gaagacagct atgggcagca gtggacctac gagcagagga aaatcgtgga
     8341 gttcacttgc cacacagcct tcttcgtcag tattgtggtg gtgcagtggg ccgacttgat
     8401 catctgcaag acccggagga attccgtttt ccagcagggg atgaagaaca agatcttaat
     8461 atttggcctc tttgaagaga cagcccttgc tgctttcctt tcctactgcc ctggaatggg
     8521 tgttgccctt aggatgtatc ccctcaaacc tacctggtgg ttctgtgcct tcccctactc
     8581 cctgctcatc ttcgtgtacg acgaagtcag gaagctcatc atcaggcggc gccctggcgg
     8641 ctgggtggag aaggaaacct actactaggc ggccgcctgg gcctcatggg ccttccgctc
     8701 actgcccgct ttccagtgcg gccgctttcg aatctagagc ctgcagtctc gacaagcttg
     8761 tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     8821 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     8881 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     8941 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     9001 cttatcatgt ctggatctga tcactgcttg agcctaggag atccgaacca gataagtgaa
     9061 atctagttcc aaactatttt gtcattttta attttcgtat tagcttacga cgctacaccc
     9121 agttcccatc tattttgtca ctcttcccta aataatcctt aaaaactcca tttccacccc
     9181 tcccagttcc caactatttt gtccgcccac agcggggcat ttttcttcct gttatgtttt
     9241 taatcaaaca tcctgccaac tccatgtgac aaaccgtcat cttcggctac ttt
//
