LOCUS       sequence-374306-        9306 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 9306)
  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 9306)
  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: MON +N122H
FEATURES             Location/Qualifiers
     source          1..9306
                     /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    8896..9030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8933..8952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8987..9006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9059..9224
                     /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 gctttttatg tcaaatttca tatcaaaacg
     4321 tcccatgaaa cgatccttgt cactgtagtg gatgttctga gcataggcct tgcattcgat
     4381 acgcaactcc atgtcgtagg tcaggttggt gaactgcacg gccaaaaggg gctggagata
     4441 cttggcctgg aggagcttgc cataatacgg gtagtattgt agaggaaatc cgccatagcc
     4501 acccatgcca tagtattgca cagggccaat tttgtcgaga tcttcctctt tcttagcagt
     4561 acaatagaca ggaatgatat tagcattgta ttttgcttgc aggtcagccg gaagggaatc
     4621 atttgctgga ggctttggta tgaagcccag aatcctgttg agtttgatga ggacacaggg
     4681 ctttccatct ttatagccat aataagggtc atttattccg gagcaatttt ccagccaggc
     4741 gcgattgaac ttgcaagatg gtagcttgcc tctagaggag tcataggcac ctctgtcttt
     4801 ggatgcagaa ggtgtttccc cacaggactc aaactccata tctgtatctt tctgctgtga
     4861 tatattataa ggtacaagaa actggtcgag tagatttgtg tatttcttgt aggtgctggg
     4921 atcagaaggg ttgaaagaaa tctccatttt aagcacttgg ggaacatttg tcagtcctgg
     4981 gggtgcaact cggtcctgat acttgggctg gaattcgctg atggtgagca gcatgacttg
     5041 aatggtccca atgaagatac ctgccaggca gccgtagaag atcacataga agaggaggat
     5101 cttaaaccag ctgccgccgg tcctccccag gaattccttc ttctccgagt tccagatgaa
     5161 tttcttccag ttgccatccc cttcctttgc ttttccgcga 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 aaggcctagg cgcgccagtc gacatgggca aagggactgg acgtgacaag tatgagcctg
     5641 cagctacatc tgagcatgga gccaagaaga aaaaggggaa gaaagaaaag gatatggatg
     5701 aactgaaaaa agaagttgca ctggatgacc acaaactcag ccttgatgag cttcatcgaa
     5761 aatatggaac ggatctcagc cggggtctga ctcctgcacg ggctgcagag atactggctc
     5821 gtgatggccc caatgctctc acccctcctc ccacgacccc agaatgggtt aaattctgca
     5881 ggcagctctt tggaggcttc tccctgttgc tgtggatagg atctattctt tgttttctgg
     5941 catacggtat tacagctgca actgaagagg agggaaataa cgatcatctg tatctgggtg
     6001 ttgtgttggc cgctgttgtc atcataactg ggtgcttctc atactaccaa gaagcaaaga
     6061 gttcaaaaat catggagtcc ttcaagaaca tggtgcctca gcaagctctg gtgatacgaa
     6121 atggagagaa gttgagcatc aatgctgaag gggtggttgt tggggaccta gtggaagtga
     6181 agggaggaga cagaatccca gctgacctga ggatcatttc agcccatggc tgcaaggtgg
     6241 ataactcttc tctcacgggt gaatcagaac ctcagacaag gtctccagac ttcacaaatg
     6301 aaaacccact ggagaccaga aatatcgctt tcttctctac caactgtgtt gaaggtactg
     6361 cacgtggtgt agtcattaat actggtgacc gcactgtgat gggccgtatt gcttcattgg
     6421 catctggact ggaaggggga cagacaccca tttctataga gatcgagcat tttatccata
     6481 taattactgg ggtggctgtg ttccttggcg tctccttctt catcctttcc ctgatcctgg
     6541 agtacacttg gctggaggct gttatctttc tgattggcat cattgtggcc aacgttcctg
     6601 agggtctgct tgctacagtc acagtatgtc tgaccctaac agccaagcgc atggctcgca
     6661 aaaattgtct ggtgaagaat ttggaagccg tagaaaccct cggctccacc tccacaatct
     6721 gctctgacaa aacaggcacc ctgactcaga accgcatgac agttgcccac atgtggtttg
     6781 acaaccagat ccacgaagct gacactacag aaaaccagag cggtgcctct tttgacaaga
     6841 catctcccac atggactgct ttatccagaa ttgctggact ttgcaaccgg gcagtgtttc
     6901 aggctaacca agaaaatgtt cccattctga agagggcagt ggcaggtgat gcctctgaat
     6961 cagcactgct gaaatgcatt gaactctgct gtggatctgt caaggacctg cgggagaaga
     7021 acaccaaggt ggtggagata ccattcaact ctaccaacaa ataccaactg tcaatccaca
     7081 agaatgcaaa cccggcagaa tcacggtacc tgcttgtgat gaaaggagcc ccagagagaa
     7141 tcctagaccg ctgcagcacc atcctgatgc atggaaaaga gcaaccctta gatgaagagg
     7201 ccagagaagc atttcagaat gcctaccttg aattaggtgg cctgggagag agagtactgg
     7261 gtttttgcca cttggccctg ccagacgaac agttccctga agatttccag tttgatacag
     7321 atgatgtaaa cttcccagtg gacaagctct gttttgtggg attgatttct atgattgacc
     7381 ctcctcgtgc ggctgtgcct gatgctgttg gcaaatgcag aagtgctgga ataaaggtta
     7441 ttatggtgac aggagaccat cccatcacag ctaaagccat tgccaaagga gttggcatca
     7501 tttctgaggg caacgagact gtggaagata ttgctcttcg ccttaacatt cctgtcaacc
     7561 aggtcaaccc cagggatgca aaagcttgtg tgattcatgg ctctgacttg aaggacatga
     7621 acagtgagca gctagacgat atcctgaaac atcatacaga aatagtcttt gccagaactt
     7681 ctcctcagca gaagcttatc attgtggaag gctgccaacg acagggtgct attgtagccg
     7741 tcacaggtga tggtgtgaat gattctcctg ctctgaagaa ggcagacatt ggtgttgcta
     7801 tgggcattgc tgggtcagat gtctccaaac aagcagctga catgattctg ctggatgaca
     7861 actttgcctc tattgtgact ggggttgaag agggtcgtct gatttttgat aacctgaaga
     7921 aatcaattgc ttataccttg accagtaaca tccctgagat tacacctttc ctcatcttta
     7981 taattgccaa catccctcta ccactcggga cagtcaccat cttgtgcatt gacttgggca
     8041 ccgacatggt tcctgccatc tctctggcat atgagcaagc agaaagtgac atcatgaagc
     8101 gacagcccag aaatcccaag acggacaaac tcgtaaacga aaggctaatc agcatggctt
     8161 atgggcagat tggaatgatt caagcactgg gaggcttctt cacatatttt gtgatcctgg
     8221 cagaaaacgg cttcttgcca tcatccttgt tagggattag agtggcatgg gatgaccgat
     8281 ggattaatga tgtagaagac agttatggac agcagtggac atatgagcaa aggaaaatcg
     8341 tggaattcac ttgtcacaca tccttctttg tcagcattgt ggtggtgcag tgggcagact
     8401 tggttatctg taagaccaga aggaattctg tctttcagca aggaatgaag aacaagatcc
     8461 tgatatttgg tctctttgaa gagactgccc tagctgcctt cctgtcttac tgccctggaa
     8521 tggatgttgc cttgaggatg tatcctctca agccgacttg gtggttctgt gccttcccct
     8581 actctcttct catctttgtg tatgatgaag taagaaaact cattcttaga cgtagcccag
     8641 gaggctgggt ggaaaaggaa acctactact aagcggccgc ttttaattaa ctggcctcat
     8701 gggccttccg ctcactgccc gctttccagt gcggccgctt tcgaatctag agcctgcagt
     8761 ctcgacaagc ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag
     8821 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     8881 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     8941 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9001 ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa
     9061 ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta
     9121 cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact
     9181 ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt
     9241 cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc
     9301 tacttt
//
