LOCUS       sequence-374324-        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: TEG +H122D
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(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    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 ggttctaatc tcaaatttca tatcaaaacg
     4321 tccctggaaa cgatctttgt cactgtagtg gatattctga ccataggcct tgcattcgat
     4381 gcgcacctcc acgtcagttg tcaggttggt gaactgcact gctataaggg gctggaggta
     4441 tttaggctgg aggagcttgc cgtaatatgg gtagtactgt agagggaagc cgccgaattt
     4501 acccaagcca tagtattcca ctgggccaat tttgtcaaca tcttcctctt tctttgctgt
     4561 gcagagaatt ggaataatat agctgttata agttgcctgc agctcagatg ggagggaatc
     4621 atttgaagga ggcttaggga cgaagcccag aatcctgttg agtttgatga tgacacatgg
     4681 cttgccaagt tggtaaccaa atgtagggtc ctgtaaccca gagcagttgt ctagccactc
     4741 gcggaagaac ctgcaggaag taatcttgcc tagagaggga tcggaatctt tttctctggg
     4801 ggcagaaggc tgagtgtcac aagtctcaaa ctccatgtct ttttcatcct gttgatcttt
     4861 attgtaaggc ttcaggaaac tgtcaagcat ctttacgtat ttttggtatg tgctggggtc
     4921 attggggttg aaggaaattt ccattttaag tacctggggg atatttgtca atcctggggg
     4981 tgccactcgg tcttgatatt tgggccggaa ttcgctgaca gtgagcagca tgacttggat
     5041 ggtcccgatg aagatgccag ccagacagcc atagaagatc acgtagaaga gaaggatctt
     5101 aaaccaactg cctccggttc tcccgagaaa ttccttcttc tctgagttcc agataaattt
     5161 cttccaattg ccatcccctt ccttcgcttt cccgcgagcc 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 aagggactgg acgtgacaag tatgagcctg cagctacatc
     5641 tgagcatgga gccaaaaaga aaaaggggaa gaaggaaaag gatatggatg aactgaaaaa
     5701 ggaagttcaa ctggatgatc acaaactcag ccttgacgaa ctccatcaca aatatggaac
     5761 agacctcagc cggggtctga ctcctgcaag ggctgctgag attttggctc gtgatggtcc
     5821 caatgctctc actccacctc ccacaacacc agagtgggtt aaattctgtc ggcagctatt
     5881 tggtggtttc tctcttctct tatggattgg ggcaatcctt tgtttgctgg catatggaat
     5941 tcaagctgca gcggatgggg aacctaacaa tgatgatctg tatctgggtg ttgtgcttgc
     6001 tgctgtagtc atcataactg gttgtttttc ttactaccaa gaagcaaaga gttcaaagat
     6061 catggaatcc ttcaagaaca tggtgcctca acaagcactt gtgatccgaa atggggaaaa
     6121 aatgagcatc aatgctgaag gggtggtcgt tggagacctg gtggaggtaa aaggaggaga
     6181 cagaatccca gctgacctga ggatcatttc agctcatggc tgcaaggtgg acaattcttc
     6241 cctcacgggt gaatcagagc ctcagacaag atctccagat ttcaccaatg agaaccctct
     6301 ggaaacgaga aatattgctt tcttctctac taactgtgtt gaaggaacag cccgtgggat
     6361 cgtcattaac actggtgatc gcactgtaat gggtcgcatt gctaccctgg cctcgggatt
     6421 ggaaggcggg cagacgccta ttgctgtgga gattgagcac tttatccaca tcattactgg
     6481 agttgctgta ttccttggtg tttccttctt catcctctca ctgatccttg agtacacttg
     6541 gttggaggct gttatctttc ttattggtat cattgttgcc aacgtcccag aaggtctgct
     6601 tgctacagtc acagtatgct tgacactaac tgccaagcgc atggctcgta agaattgcct
     6661 tgtgaagaat ttggaagctg tagaaaccct gggctccaca tctacaattt gctctgacaa
     6721 aacagggacc ttgactcaga accgcatgac agttgctcac atgtggtttg ataaccagat
     6781 tcatgaggct gatactacag aaaaccagag cggtgcttcc tttgacaaga catctcccac
     6841 ttggactgct ttagccagaa ttgctggact ctgtaaccgt gctgtgttcc aggctaacca
     6901 agataatgtt cctattctga agagggcagt ggcaggtgat gcttctgagt ctgcactcct
     6961 caaatgcatt gaactgtgct gcggatctgt caaggaatta agggagaaaa acccgaaagt
     7021 ggtcgagata ccattcaact caaccaacaa ataccagctg tccatccaca ataatgcaaa
     7081 ctcatcagaa tcacgatacc tacttgtgat gaaaggagcc ccagagagaa tcctggaccg
     7141 ttgcagtact atcctgatgc atggaaaaga gcaaccctta gatgaagagg caaaagatgc
     7201 atttcagaat gcctacctag aattgggagg cctaggggag agagtcttag gcttttgcca
     7261 cctggccctg cctgatgatc agttccccga aggcttcgag tttgacacag atgatgtgaa
     7321 cttcccagta gagaagctct gtttcgttgg gctgatttca atgattgacc ctccccgtgc
     7381 tgctgtgcct gatgctgttg gcaaatgcag gagtgccgga ataaaggtta ttatggtgac
     7441 tggagaccat ccaatcacag ctaaagctat tgccaaggga gttggcataa tctctgaagg
     7501 caacgagact gtagaagaca tcgctcttcg actcaacatt cctgtcagcc aggtcaatcc
     7561 cagggatgcc aaagcctgtg ttattcatgg atctgatttg aaggacatga caagtgagca
     7621 gctagatgac attctgaggc accatacaga aatagttttt gccaggacat ctcctcaaca
     7681 gaagcttata attgtggaag gttgccagcg acagggtgct attgtagctg tcacaggtga
     7741 tggtgtcaat gattcccctg ctttaaagaa agcagatatc ggcgttgcta tgggcattgc
     7801 tggatctgat gtctccaaac aggcagctga catgattctg ctggatgaca actttgcctc
     7861 tattgttact ggagttgaag agggtcgcct gatttttgac aacctgaaga agtcaattgc
     7921 ttacactttg actagtaaca tccctgagat aacacctttt ctgatcttca taattgccaa
     7981 catacctctg ccactgggta cagtcaccat cctctgcatt gatttgggca ctgatatggt
     8041 tcctgccatc tctctggcat atgagcaagc agaaagtgac atcatgaaga ggcagcccag
     8101 aaatcccaag actgacaaac tagtaaatga aaggttgatc agcatggctt acggtcaaat
     8161 cggaatgatc caagcattgg gaggcttctt tgcatatttc gtaatccttg cagagaatgg
     8221 cttcttgcca tcttctctgt tagggatcag agtggcatgg gatgaccgat ggattaatga
     8281 tgtagaggac agttatggac aacagtggac ttatgaacaa agaaagattg tggaattcac
     8341 atgtcacacg gcctttttcg ttagcattgt ggtggtgcag tgggcagact tgattatctg
     8401 taagaccagg aggaattctg tcttccagca aggaatgaag aacaaaatct taatatttgg
     8461 tctctttgaa gagactgccc tggctgcctt cctctcatat tgccctggaa tggatgttgc
     8521 tttgaggatg tatcctctca agccaacttg gtggttttgt gccttcccct actctctcct
     8581 catttttgtg tatgacgaag tgagaaaact cattctcaga cgcagccctg gaggctgggt
     8641 ggaaaaggaa acgtactact aagcggccgc 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
//
