LOCUS       sequence-374308-        9290 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 9290)
  AUTHORS   Mohammadi S, Herrera-Alvarez S, Yang L, del Pilar Rodriguez-Ordonez 
            M, Zhang K, Storz JF, Dobler S, Crawford AJ, Andolfatto P
  TITLE     Constraints on the evolution of toxin-resistant Na,K-ATPases have 
            limited dependence on sequence divergence
  JOURNAL   bioRxiv 2021.11.29.470343
REFERENCE   2  (bases 1 to 9290)
  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: "bioRxiv 
            2021.11.29.470343"
COMMENT     Sequence Label: SNG +R111Q
FEATURES             Location/Qualifiers
     source          1..9290
                     /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    8880..9014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8917..8936)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8971..8990
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9043..9208
                     /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 gtggaccagg aggcggccga caatgataat ctgtacttgg gtattgtgtt
     6001 ggcagctgtg gttatcatta ctggctgttt ctcttattac caagaagcaa aaagttccaa
     6061 gatcatggag tccttcaaga acatggtgcc tcagcaagct cttgtagtca gaaatggtga
     6121 gaagatgagc ataaatgctg aaggtgttgt agttggagat ctagtggagg tgaaaggagg
     6181 agacagaatt ccagccgacc ttcggatcat atctgcacat ggttgcaagg tagataactc
     6241 ctcacttact ggtgaatcag agcctcagac cagatctcca gacttctcca atgagaatcc
     6301 cctggagacc aggaacattg ccttcttttc caccaactgt gtggaaggca ctgcccgtgg
     6361 cattgtcatt agcactgggg atcgcactgt aatgggccgt attgccagtc tggcttctgg
     6421 gctggaaggg gggaaaactc caattgccat ggagattgag cactttatcc acctcatcac
     6481 tggagtagct gtcttcctgg gtgtgacttt cttcatcctt tcactcatcc ttgagtacac
     6541 atggctggag gctgtaatct tcctcatcgg gatcattgtt gccaatgtcc ctgaagggct
     6601 gcttgcaaca gtcacggtat gtctgacact aacagccaag cgtatggctc gtaagaactg
     6661 cttggtgaag aacctcgaag ctgtggaaac cctgggttcc acgtccacta tctgttctga
     6721 caaaacaggc acgctgacac agaatcgcat gacggttgcg cacatgtggt ttgacaatca
     6781 gattcatgag gctgatacta cagagaacca gagtggtgct tcctttgaca agagctcagc
     6841 tacttggact gctttgtcca gagttgcagg tctctgcaac cgtgctgtgt ttcaggccaa
     6901 ccaggaaaat gtaccaatac ttaagagagc agtggcggga gatgcctctg agtctgcact
     6961 tctgaaatgc attgaattgt gctgtggtcc tgtcaaggag atgagagaaa ggtatcccaa
     7021 agtggtggaa ataccattta actctaccaa caagtatcag ctgtctatcc acaaaaatgc
     7081 aaatccgtca gaatcccgtt acttactggt gatgaaggga gctccagaga ggattttgga
     7141 tcgctgcagc accattctta ttcatggcaa agagcaacca ctggatgagg aaatgaaaga
     7201 ttcttttcaa aatgcctacc ttgagttggg aggccttggg gagagagtgt taggattctg
     7261 tcacttggct ctgcctgatg atcagttccc tgatggcttc cagtttgata cagacgaagt
     7321 gaactttcct gtagaaaaac tctgctttgt aggactgatg tctatgattg accctcctcg
     7381 tgctgctgtg ccagatgctg ttggcaaatg cagaagcgct gggatcaagg ttatcatggt
     7441 tacgggagac catccgatca cagccaaagc cattgccaag ggtgtcggca tcatctctga
     7501 gggcaatgaa acagttgaag atattgctgc tcgactcaac attcctgtca gccaggtcaa
     7561 ccctagggat gccaaagctt gtgttgttca tggctcagat ttgaaggaca tgactagtga
     7621 gcaactggat gacatcttgc ttcatcatac tgaaattgtc tttgccagga catctcctca
     7681 gcagaagctt ataattgtgg aaggctgtca gcgacagggt gccattgtag cagtcacagg
     7741 tgatggtgtg aatgattccc cagccctgaa gaaggctgac attggtgttg ctatgggtat
     7801 tgctggctca gatgtctcca agcaggcagc tgacatgatt ctgctggatg ataactttgc
     7861 ctccattgtc actggggttg aagaagggcg tctgatcttt gataacctga agaagtctat
     7921 tgcttacacc ttgaccagta acattcctga aatcacgccg ttcctgatct tcatcattgc
     7981 aaacataccc cttccactgg gaacagtcac catcctctgc attgacttgg gcactgacat
     8041 ggtccctgct atctccctgg catatgagca agcagagagt gacatcatga agaggcagcc
     8101 cagaaatccc aaaacagaca agctggttaa tgaacggctg atcagcatgg cctatgggca
     8161 gattggtatg atccaggccc ttggaggttt cttcacctat tttgtaatca tggcagagaa
     8221 tgggttctgg ccttctggct tgctagggat cagagttcag tgggatgacc gatgggttaa
     8281 cgatgtggaa gacagctatg ggcagcagtg gacctacgaa cagaggaaaa tagtggagtt
     8341 cacttgccat acagccttct ttgtcagcat tgtggttgtg cagtgggcag acttgatcat
     8401 ttgtaagacc agaagaaact ctgtcttcca gcagggaatg aagaacaaga tcttaatatt
     8461 tgggctcttt gaggagactg ctctggctgc cttcctctcc tactgccctg gaatggatgt
     8521 tgctctaagg atgtaccctc tcaagccaac ctggtggttc tgcgctttcc catactctct
     8581 tctcatattc ctgtatgatg aaatcagaaa gctcattatc agacgcaacc ctggtggttg
     8641 ggtggaaaaa gagacctact actaagcggc cgcctgggcc tcatgggcct tccgctcact
     8701 gcccgctttc cagtgcggcc gctttcgaat ctagagcctg cagtctcgac aagcttgtcg
     8761 agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     8821 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     8881 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     8941 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9001 atcatgtctg gatctgatca ctgcttgagc ctaggagatc cgaaccagat aagtgaaatc
     9061 tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt
     9121 tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc
     9181 cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa
     9241 tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt
//
