LOCUS       sequence-374307-        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: FER +H122D
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 cactgtagtc aatattttga ccataggcct tgcactcgat
     4381 acgcagctcc atatcagtag tcagattagt gaactgtact gctacaaggg gttgaaggta
     4441 ttttggctga atgagcttgc cataatatgg gtagtactgc agaggaaagc caccaaagcc
     4501 attgaaacca tagtattcca cagcgccaat tttgttcgac tcctcttctc tcttagcagt
     4561 gcaatgaata ggaatgacat tgggattata ttttgccagc agctcttgtg ggatggtatc
     4621 attttgtgga atcttgggtg tgtaacccag gattctgttg agtttgatga tgacacatgg
     4681 tttgccttcc tcgtaaccga aattagaatc agttatgcca gagcaatttc ccagccaact
     4741 gcgaaagaac ttgcaagatg gcagggcacc tagagagcca tcaaaaggtc ctctgtcttt
     4801 gggaggagaa ggttcagtgc cacaattctc aaaattgacc tcttgctgat tagcatcgtc
     4861 ataggccttt aagaattttt caatttgggt tacatatggc ttgtatgagt ttggatcaga
     4921 agggttgaaa gaaatctcca ttttaaggac ttggggaaca tttgacaatc ctgggggtgc
     4981 tattcggtcc tgatacttgg gctggaattc gctgatggtg agcagcagga cttgaatggt
     5041 accaatgaag atgcctgcca ggcagccata gaagatcaca tagaagagga ggattttaaa
     5101 ccaactgccg ccggtcctcc ccaaaaattc cttcttctcc gagttccaga tgaatttctt
     5161 ccagttgccc 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 gagcctgcag ctacatctga
     5641 gaatgcagca aagaagaaaa aaggaaaaaa agaaaaggat ttggatgaac taaaaaaaga
     5701 agtttcactg gatgatcaca aactcagcct tgatgaactt catcgcaaat atggaactga
     5761 tctgagtcgg ggactgtctg tggcgcgggc tgcagaaatc ctggctcgag atggccctaa
     5821 tgctctcact ccccctccca caaccccgga atgggttaaa ttctgccggc agctctttgg
     5881 aggcttctcc cttctgcttt ggattggagc tcttctttgt tttctggcgt atggtattca
     5941 tattgcaaca ggagaagagc ctaataatga cgatttatat ctaggtgttg tgctgtctgc
     6001 tgttgtcatc ataactgggt gcttctctta ctaccaagaa gcaaagagtt caaagatcat
     6061 ggagtccttc aaaaacatgg tgcctcagca agctttggta atccgaaatg gggagaaact
     6121 gagcatcaat gcagaaggcg tagctgttgg agacctggtg gaggtgaagg gtggagacag
     6181 aatcccagct gatttgagaa taatttcggc taatggatgt aaggtggaca attcatctct
     6241 cacaggtgaa tcagaacctc agacaaggtc tccagattgt acaagtgata acccactaga
     6301 gaccagaaat attgcatttt tttccactaa ttgtgttgaa ggtactgccc gtggtatagt
     6361 catcaacact ggtgaccgaa ccgtgatggg ccgcattgct acgttggcat ctgggttgca
     6421 aggaggaaag actcccattg ctgtggagat tgagcatttt atccacataa ttactagtgt
     6481 ggccgtgttc ctgggtgttt ccttttttat cctctcattg atcctcgagt acacttggct
     6541 agaagctgtg atattcctta tcggcatcat tgttgccaat gtccctgagg gtctgcttgc
     6601 tacagtgaca gtatgtctga cactcacagc caagcgcatg gctcgtaaga attgtcttgt
     6661 gaagaactta gaagccgtgg agactctggg ctctacttct accatctgct ctgacaaaac
     6721 aggcaccctg actcagaacc gcatgactgt cgcccacatg tggttcgata accagatcca
     6781 tgaagctgac acaacagaaa accagagtgg tgcttctttt gacagaacat ctccaacatg
     6841 gactgcttta gccaagattg cagggctatg taaccgggcc gtatttcagg ctaaccaaga
     6901 aaatgttccc attttgaaga gagcagtggc aggagacgct tcagaatctg cactcctaaa
     6961 atgcattgaa ctttgctgtg gatctgtcaa ggaactgagg gagaagaacc ccaaagtagt
     7021 ggagatccca ttcaactcta ccaataaata ccaactgtca attcacaaga atccaaactc
     7081 gtcagaatca cgatatctgt tggtgatgaa aggtgcaccg gagagaattc tcgaccgttg
     7141 cagttccatc ctgatacacg gaaaagaagt tcccttggat gaggaggcca aagatgcatt
     7201 tcagaatgcc tatcttgaat tgggaggcct gggggagaga gttctgggct tctgccacct
     7261 gtctctgcct gatgaacagt ttcctgaaga cttccaattc gatacagatg atgtgaactt
     7321 ccctgttgat aagctgtgtt ttgttggttt aatttctatg attgaccctc ctcgtgctgc
     7381 tgtgcctgat gctgttggca aatgcagaag tgctggaata aaggtcatta tggtgacagg
     7441 agaccacccc atcacagcaa aagctattgc caagggtgta ggcatcatct ccgagggaaa
     7501 cgagactgtg gaagatattg ctcttcgcct caacattcct atcaaccagg tcaaccccag
     7561 agatgccaaa gcctgtgtga ttcatgggac tgacttgaag gatatgagca gtgaacaact
     7621 agatgatatt ctgagacatc atacagaaat agtctttgct agaacatctc cacaacagaa
     7681 gcttatcatt gtagaaggct gccaacggca gggtgctatt gtagctgtca ctggagatgg
     7741 tgtgaacgat tcccctgctt tgaaaaaggc agacattggt gttgctatgg gcattgctgg
     7801 gtcagatgtc tctaaacagg cagctgacat gattctactg gatgacaact ttgcttctat
     7861 tgttactgga gttgaagaag gccgcctgat ttttgataat ctgaagaaat caattgccta
     7921 tactttgacc agtaacattc ctgagattac gcctttcctt atcttcataa tcgccaacat
     7981 ccctctaccg ctggggacag tcaccatctt atgcattgac ttaggcactg atatggttcc
     8041 tgcaatatct ctggcatatg agcaagcaga aagtgacatc atgaagagac agcccagaaa
     8101 ctcaaagaca gacaagctgg tgaacgaaag gctaattagc atggcttatg gtcagattgg
     8161 gatgatccaa gccttgggtg gcttcttcac atactttgta atcctcgcag agaacggctt
     8221 cttgccatcc agtttactgg ggattagagt gttgtgggat gaccggtggg taaatgatgt
     8281 ggaggacagc tatggacagc aatggactta tgagcaaagg aaaattgtag aattcacttg
     8341 ccacacagca ttctttgtca gcattgtggt ggtacagtgg gcagacttga ttatctgtaa
     8401 gaccagaagg aattctgtct tccagcaagg aatgaaaaac aagatcctaa tatttggtct
     8461 ctttgaagaa actgctcttg ctgccttcct ctcttattgc ccaggaatgg gtgttgcctt
     8521 gagaatgtat ccattgaaac cttcatggtg gttttgcgct ttcccttatt ctctcctcat
     8581 ctttgtctat gatgaagtaa gaaaattaat catcagacgt agccctggag gctgggtgga
     8641 aaaagaaacc tattactaag 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
//
