LOCUS       sequence-374311-        9284 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 9284)
  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 9284)
  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: KEE +E111H
FEATURES             Location/Qualifiers
     source          1..9284
                     /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    8874..9008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8911..8930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8965..8984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9037..9202
                     /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 gctttttatc tcaattttca tatcaaaacg
     4321 tccctggaaa cgatctttgt cactgtagcc aatattttca ccataggcct tgcactcgat
     4381 acgcagctcc atatcggtgg tcaaattagt gaaccgtact gctacaaggg gttgaaggta
     4441 tttctgctga atgagcttgc cataatatgg gtagtattgt agaggaaagc caccatagcc
     4501 acccaaacca aagtattcca cggggccaat tttgctcaaa tcctctgatt tcttagcagt
     4561 gcaatgaatg ggaatgacat tgggattata ttttgccagc acctcaggtg ggatggaatc
     4621 attttgtgga agcttaggta tgtaacccag aactctgttg agtttgatga tgacacatgg
     4681 tttgccatct tcgtaaccat aattaggatc gtctatgcca gagcaatttc ccagccaact
     4741 gcgaaagaac ttgcaggatg gcatggcacc ttgtgtgcca tcaaaaggtc ctctctcttt
     4801 gggaggagaa ggttcagtcc cacaattctc aaaattgatg tcttgctgat taacactttc
     4861 atagcccttt aagaaatctt caatttggtt tatatatggc ttgtatgttt ttggatcaga
     4921 aggggtgaaa gaaatctcca ttttaaggac ttggggaaca tttgtcaatc ctgggggcgc
     4981 tattcggtcc tgatacttgg gctggaattc gctgacagtg agcagcatga cttgaatggt
     5041 cccaatgaag atgcctgcca ggcagccata gaagatcacg tagaagagga ggatcttaaa
     5101 ccaactgccg ccggtcctcc ctaaaaattc cttcttctcc gagttccaga tgaatttctt
     5161 ccagctgcca tccccttcct tggctttccc gcgagccatc 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 atgagcaaag ggactggacg tgacaagtat gaacctgcag ctacatctga
     5641 gaatgcagca aagaagaaaa aaggaaaaaa agaaaaggat atggatgaac taaaaaaaga
     5701 agtttcactg gatgatcaca aactcagcct tgatgaactt catcgtaaat atggaactga
     5761 tctcagccgg ggactgtctg ctcagcgtgc tgcagaaatc ttggctcgag atggccctaa
     5821 tgctctcact ccccctccca caaccccaga atggattaaa ttctgccgac aactgtttgg
     5881 aggtttcgcc cttctgcttt ggattggagc ttttctttgt tttctgttgt ttggtattca
     5941 ttctgcaaca ggagagaaac ctaataatga caatttatat ctaggtattg tgctgtctgc
     6001 tgttgtcatc ataactgggt gtttctctta ctaccaagaa gcaaagagtt caaagatcat
     6061 ggagtccttc aaaaacatgg tgcctcagca agcgttggta gtccgaaatg gggagaaact
     6121 gagcatcaat gctgaaggtg tagctgttgg agacctagtg gaagtgaagg gaggagacag
     6181 aatcccagct gatttgagaa taatttcagc taatggatgt aaggtggaca attcatctct
     6241 cacaggtgaa tcagagcctc agacaaggtc tccagattgt acaaatgata acccactaga
     6301 gaccagaaat attgcttttt tttccaccaa ctgtgtggaa ggcactgccc gtggtatagt
     6361 tatcaacact ggtgaccgaa ctgtgatggg ccgcattgct acgttggctt ctgggttgga
     6421 aggtggacag actcccattg ctgcggagat tgagcatttt atccacataa ttactggtgt
     6481 ggccgtgttc ctgggtgttt ccttttttat cctctcattg atcctcaagt acacttggct
     6541 agaagctgta atatttctta tcggcatcat tgttgccaat gtcccggagg gtctgcttgc
     6601 tacagtgaca gtatgtctga cactcacagc caagcgcatg gctcgtaaga attgccttgt
     6661 gaagaactta gaagccgtgg agactctggg ctctacttct acaatctgtt ctgacaaaac
     6721 aggcaccctg acccagaacc gcatgaccgt tgcccacatg tggttcgata accagatcca
     6781 tgaagctgac accacagaaa accagagtgg tgcttctttt gacaaaacat ctccaacatg
     6841 ggctgcttta gccaagattg caggactatg taaccgggct gtatttcaag ctaaccaaga
     6901 aaatgttccc attttgaaga gagcagtggc cggagatgct tctgaatctg ccctcctaaa
     6961 atgcattgaa ctttgctgtg gatctgtcaa ggaactgagg gagaagaacc ccaaagtagt
     7021 ggagattccg ttcaactcta ccaataaata ccaactgtca attcacaaga atccaaacac
     7081 gtcagaatca cgctatctgt tggtgatgaa aggtgcaccg gagagaattc tcgatcgttg
     7141 cagttccatc ctgatgcacg gaaaagaact tcccttggat gaggaggcca aagatgcatt
     7201 tcagaatgcc tatcttgaat tgggaggctt gggggagaga gttctgggct tctgtcacct
     7261 ggctctgcct gatgaacagt ttcctgaagg cttccaattt gatacagatg atgtgaactt
     7321 ccctgtagag aagctgtgtt ttgtgggttt aatttctatg attgatcctc ctcgtgctgc
     7381 tgtgcctgat gctgttggca aatgcagaag tgctggaata aaggtgatta tggtgacagg
     7441 agaccacccc atcacagcga aagctattgc caagggtgta ggcatcatct ctgagggaaa
     7501 tgagaccgtg gaagatattg ctcttcgcct caacatccct gtcaaccagg tcaaccccag
     7561 agatgccaaa gcctgtgtga ttcatgggtc tgacttgaag gatatgacca gtgaacaact
     7621 agatgatatt ctgaaacacc atacagaaat agtctttgct agaacgtctc cacaacagaa
     7681 gcttatcatt gtagaaggct gccaacggca gggtgctatt gtagctgtca ctggagatgg
     7741 cgtgaatgat tcccctgctt tgaaaaaggc agacattggt gttgctatgg gcattgctgg
     7801 gtcagatgtc tctaaacagg cagctgacat gattctgctg gatgacaact ttgcttctat
     7861 tgttactgga gttgaagaag gtcgcctgat ttttgataat ctgaagaaat caattgccta
     7921 tactttgacc agtaacattc ctgagattac gcctttcctt atcttcataa ttgccaacat
     7981 ccccctaccg ctggggacag tcaccatctt atgtattgac ttaggcactg atatggttcc
     8041 tgcaatttct ttggcatatg agcaagcaga aagcgacatc atgaagaggc agcccagaaa
     8101 ccccaataca gacaaactgg taaacgaaag gctgattagc atggcatatg gtcagattgg
     8161 gatgatccaa gccttgggtg gcttcttcac atactttgta atcctcgcag agaatggctt
     8221 cttgccatcc tctttactgg ggattagagt gatgtgggat gatcgatggg taaatgatgt
     8281 ggaggacagc tatggacagc agtggactta tgagcaaagg aaaattgtgg aattcacttg
     8341 ccacacatca ttctttgtca gcattgtggt ggtgcagtgg gcagacttgg ttatctgtaa
     8401 gaccagaagg aattctgtct tccagcaagg aatgaagaac aagatcctaa tatttggtct
     8461 ctttgaagaa actgctcttg ctgccttcct ctcttactgc ccaggaatgg ccgttgcctt
     8521 gagaatgtac ccattgaaac cttcatggtg gttttgtgct ttcccttatt ctctcctcat
     8581 ctttgtctat gatgaagtaa gaaaactaat catcaggcgc agccctggag gctgggtgga
     8641 aagagaaacc tactactaag cggccgcctg ggcctcatgg gccttccgct cactgcccgc
     8701 tttccagtgc ggccgctttc gaatctagag cctgcagtct cgacaagctt gtcgagaagt
     8761 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     8821 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     8881 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8941 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     9001 tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga aatctagttc
     9061 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
     9121 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
     9181 ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac
     9241 atcctgccaa ctccatgtga caaaccgtca tcttcggcta cttt
//
