LOCUS       sequence-374214-        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, 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 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: "bioRxiv 
            2021.11.29.470343"
COMMENT     Sequence Label: RAT +D122H
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
                     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(5234..5343)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5367..5458
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5383..5400
                     /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 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 gttctcacca tacgccttac actcaatgcg
     4381 gatttcagtg tccaaggtga ggttggtgaa ctgcacggcc agcaggggct gcaggtactt
     4441 gggctgcagg agtttgccgt agtagggata gtactgcaga ggaaagccat agaatccgcc
     4501 catcccaaag tactctatgt ttccaacttt atccttatcc tcatcgcgct tgccagtgca
     4561 ctggacaggt aggacgtttg gattatactt catcgtcaga gggtaagtct ccaaggattc
     4621 attcttggga ggcttaggtt tgaagcccag cactcggttg agcttgataa tgatacaggg
     4681 cttcccctct ttgtagccgt aggattcatc attgagacca gagcagttcc ccagccagtc
     4741 aagcttgaac ctgcacacct tgcgctctcc tcgttcatga ttgaactctc cccgctcctt
     4801 gggttcactg ggcatactgc cacaatcctc gaaaatcatg tcgtccttct gggccgaatc
     4861 tttgtacttt tccaggaacc tgatgatgtt tagcacatag gcctcgtagc tcttggggtc
     4921 attaggacgg aaggaaattt cagtcttttg gatctgagga atctgtgtca atcctggcgg
     4981 ggccacacgg tcctggtacg tgggtttcag ctcactgatg gtaagcagca tcacttggat
     5041 ggtcccgatg aagatgccgg ccaggcagcc atagaatatc acgtagaaca gaaggatctt
     5101 aaaccaacta ccaccggtcc tgcccaaaaa ctccttcttc tccgagttcc agatgaattt
     5161 cttccagctg ccttcctcct tggcttttcc gcgggccatc tcgagatccc gggtgatcaa
     5221 gtcttcgtcg agtgattgta aataaaatgt aatttacagt atagtatttt aattaatata
     5281 caaatgattt gataataatt cttatttaac tataatatat tgtgttgggt tgaattaaag
     5341 gtccgtatac tccggaatat taatagatca tggagataat taaaatgata accatctcgc
     5401 aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac ctataaatat
     5461 tccggattat tcataccgtc ccaccatcgg gcgcggatcc cggtccgaag cgcgcggaat
     5521 tcaaaggcct acgtcgacga gctcacctat agggcgaatt ggcggaaggc cgtcaaggcc
     5581 gcatgtcgac atggggaagg gggttggacg agacaagtat gagcccgcag ctgtatcaga
     5641 acatggggac aagaagagca agaaggcgaa gaaggaaagg gacatggacg aactcaagaa
     5701 ggaagtgtct atggacgacc ataaactcag cctggatgaa ctccatcgta aatacggaac
     5761 agatttgagc cgaggcctaa cacccgcaag ggccgctgag atcctggctc gggatggccc
     5821 caacgccctc acgccccctc ccactactcc cgagtgggtc aaattctgtc ggcagctgtt
     5881 cggtggcttc tccatgttac tgtggattgg agccattctt tgtttcttgg cttatggcat
     5941 ccgaagtgct acagaagagg aaccaccaaa tgatcatctg tacctcgggg tcgtgctgtc
     6001 tgctgtcgtc atcataactg gctgtttctc ctattatcaa gaagcaaaaa gctccaagat
     6061 catggaatcc ttcaagaaca tggtccctca gcaagccctc gtgattcgaa atggagagaa
     6121 gatgagcatc aacgcagagg atgtcgtcgt tggtgatctg gtggaggtga agggcggaga
     6181 ccgaatccct gctgatctca gaatcatatc tgcaaatggc tgcaaggtgg ataactcctc
     6241 actcactggt gaatcagaac cccagactcg gtccccggat ttcacaaacg agaacccctt
     6301 ggagacaagg aacattgcct tcttctcaac caactgtgtt gaaggaactg cacgtggcat
     6361 cgttgtgtac actggggatc gcaccgtgat gggcaggatc gccacccttg cttctgggct
     6421 ggaaggcggc cagaccccca ttgctgaaga aatcgagcac ttcatccacc tcatcacggg
     6481 tgtggccgtg ttcctggggg tgtctttctt cattctctct ctgatccttg agtacacctg
     6541 gctcgaggct gtcatcttcc tcattggtat catcgtagcc aacgtgccgg aaggtttgct
     6601 ggccaccgtc acggtatgtc tgacgctcac tgccaagcgc atggcgagga agaactgcct
     6661 ggtgaagaac ctggaagctg tggagacctt ggggtccaca tccaccatct gctccgacaa
     6721 gactggaact ctgactcaga accggatgac agtggctcac atgtggtttg acaatcaaat
     6781 ccatgaagct gacaccacag agaatcagag tggggtctcc tttgacaaga cgtcagccac
     6841 ctggttcgct ctgtccagaa ttgctggtct ctgtaacagg gcagtgtttc aggctaacca
     6901 agaaaacctg cctatcctta agcgtgcagt agcgggagat gcttccgagt cggcgctctt
     6961 aaagtgcatc gaggtctgct gtggctccgt gatggagatg agggagaagt acaccaagat
     7021 agtggagatt cctttcaact ccaccaacaa gtaccagctc tccattcaca agaacccaaa
     7081 cgcatcggag cctaagcacc tgctagtgat gaagggcgcc ccagaaagga tcctggaccg
     7141 atgcagttct atcctcctcc acggcaagga gcagcccctg gacgaagagc tgaaggacgc
     7201 ctttcagaat gcctacctag agctgggggg ccttggagag cgtgtgctag gtttctgcca
     7261 cctccttctg cctgacgaac agtttcccga aggcttccag tttgacactg atgaagtcaa
     7321 tttccccgtg gataacctct gcttcgtggg tcttatctcc atgattgacc ctcctcgagc
     7381 tgctgtcccc gatgctgtgg gcaaatgccg cagcgctggg attaaggtca tcatggtcac
     7441 aggagaccat ccaatcacag ccaaagccat tgctaagggg gtgggcatta tctcagaagg
     7501 taacgagacc gtggaagaca ttgctgcccg cctcaacatt ccagtgaacc aggtgaaccc
     7561 cagagatgcc aaggcctgtg tagtacatgg cagtgacttg aaggacatga cctctgagga
     7621 gctggatgac attttgcggt accacacgga gattgtcttt gctaggacct ctcctcaaca
     7681 gaagctcatc attgtggagg gctgccagcg gcagggtgcc atcgtggctg tcacagggga
     7741 tggtgtcaat gactctccag ctttgaaaaa ggcagatatt ggggttgcca tggggattgt
     7801 tggctcggat gtgtccaagc aagctgctga catgattctt ctggatgaca actttgcctc
     7861 catcgtgact ggagtagaag aaggtcgtct gatatttgat aacttgaaga aatccattgc
     7921 ttacacccta acaagtaaca ttccggaaat cacccccttc ttgatattta ttattgcaaa
     7981 cattccactg cccctgggca ccgtgaccat cctctgcatt gacttgggca ctgacatggt
     8041 tcccgccatc tctctggcct atgaacaggc tgaaagtgac atcatgaaga ggcagcccag
     8101 aaatcccaaa acggacaaac ttgtgaacga gcgtctgatc agcatggcct atggacagat
     8161 cggtatgatc caggccctgg gaggcttctt cacttatttt gtgattctgg ctgagaacgg
     8221 tttcctgccc tttcacctgt tgggcatccg agagacctgg gatgaccgct ggatcaatga
     8281 tgtggaggac agctacgggc agcagtggac ctacgagcag aggaagattg tggagttcac
     8341 ctgccacacg gccttctttg tcagtatcgt ggtagtgcag tgggctgact tggtcatctg
     8401 caagaccaga aggaattctg tcttccagca gggaatgaag aacaagatct taatatttgg
     8461 cctctttgaa gagacagctc ttgctgcttt cctgtcctac tgccctggga tgggtgcagc
     8521 ccttaggatg tatcccctca aacctacttg gtggttctgt gccttcccct actcccttct
     8581 catcttcgtg tatgacgagg tgcggaagct catcatcagg cgacgccctg gcggctgggt
     8641 ggagaaggaa acctactact aggcggccgc 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
//
