LOCUS       sequence-374207-        9317 bp    DNA     circular SYN 15-APR-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 9317)
  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 9317)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: OST
FEATURES             Location/Qualifiers
     source          1..9317
                     /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    8907..9041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8944..8963)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8998..9017
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9070..9235
                     /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 tcgaatttaa tatcaaaacg
     4321 tccctgaaag cggtctttgt cactgtactg gatgttctgc ccataggcct tgcattccac
     4381 acgcacttct acatcgtagg tcaggttagt gaactgcaca gccaccagcg gctgcaggta
     4441 tcgtggctgg aggagcttgc cgtagtacgg gtagtactgt agagcaaagc cagggaagcc
     4501 tcccatccca tagtactcca ctgtgcctat tttgtctgca tcttcatctc tcttggcact
     4561 gcagtggacg gggatgagat acgggctgta ttttgccata acctctggag ggagactctc
     4621 atttaccgga gccttaggtt tgaagccaat aattctgttg agcttgacta ggatgcatgg
     4681 tttgccatcc ctgtacccaa aggttggatc atttattcca gaacagtttt ccagccattc
     4741 acgtttgaat ttgcagacct tcttctggcc ttgggcatca ttatatggtc ctctgtcttt
     4801 atattcagta ggtatatccc cacagtcctg aaatattatg ttttcagttt gttgttcagc
     4861 gctgtagtcc tttaagaact tctcaaggtt tctcacgtac ggctcatagc tcttggaatc
     4921 agaggcaata aaagaaattt ctgtcttttg tacctgggga acttgagtca gtcctggagg
     4981 tgccactcga tcctgatatt tgggttcaaa ttcactgaca gtgagcaaca tcacttggat
     5041 ggtcccaatg aatatacctg ccaggcaacc atagaagatc acatagaaga ggaggatttt
     5101 aaaccagctg cccccggtgc ggcccagcag ctccttcttc tccgagttcc agatgaactt
     5161 cttccagttg ccgtcgccgt ccttggcctt cccgcgggcc 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 gccacgtgtc ttgtccgcgg taccgtcgac atgggcaagg gggttggaag agacaagtat
     5641 gagcctactg caacatccga acatggcgca aagaagaaaa aggggaagga gagggacatg
     5701 gatgagctta agaaggaagt ctttcaggat gaccacaagc tgagccttga tgaacttcat
     5761 cgtaaatatg gaacagactt gagtcgaggt ttgacacctg cacgtgcagc tgagattttg
     5821 gctcgtgatg gcccaaatgc cctcactccc ccaccaacca ctcctgaatg ggtaaagttc
     5881 tgtcggcagc tctttggagg attctcgctc ctattgtgga ttggctctat tctgtgtttt
     5941 ctggcttttg gcatacaaag tgcaatggag gaggagtcca atcatgataa tctatacctc
     6001 ggtgttgtgt tggcagctgt ggttatcatt actggctgtt tctcttacta ccaagaagca
     6061 aaaagttcca agatcatgga gtccttcaag aacatggtgc ctcagcaagc tcttgtagtc
     6121 agaaatggtg agaagatgag tataaatgct gaaggtgttg tagttggaga tctggtggag
     6181 gtgaaaggag gagacagaat tccagctgac cttcggatca tatctgctca tggttgcaag
     6241 gtagacaact cctctctcac tggggaatcg gagcctcaga ccagatctcc agacttcacc
     6301 aatgagaacc cactggagac caggaacatt gccttctttt ccaccaactg tgtagaaggc
     6361 actgctcgtg gcattgtaat tagcactggg gaccgcactg tgatgggccg tatagccagt
     6421 ctggcttctg gactggaagg agggaagact cctattgcca tggagattga gcactttatc
     6481 cacctcatca ctggggtagc tgtgttcctg ggtgtcagct tcttcatcct ctcactcatc
     6541 ctcgagtaca cttggctgga ggctgtgatc ttcctcattg ggatcattgt tgccaatgtc
     6601 cctgagggac tgcttgcaac agtcacggta tgtctgacac taacagccaa gcgtatggct
     6661 cgtaagaact gcttggtaaa gaacctggaa gctgtggaaa ctctgggctc cacatcaacc
     6721 atctgttctg acaaaacagg cactctgaca cagaaccgca tgacagttgc acacatgtgg
     6781 tttgacaatc agattcatga ggctgatact acagagaatc agagtggtgc ttcctttgac
     6841 aagagttctg ccacttggac tgccttgtcc agagttgcag gtctctgcaa ccgtgctgtg
     6901 tttcaagcca accaggaaaa tgtgcctatt cttaagaggg cagtggcagg agatgcttct
     6961 gagtctgcgc ttctgaaatg catcgaactc tgctgtggtt ctgtcaagga gatgagagaa
     7021 aggaacccca aagtggtgga aataccattt aactctacca acaagtacca gctgtctatc
     7081 cacaagaatg caaatccagc agaatctcgt tacttgctgg tgatgaaagg agccccagaa
     7141 aggatcttgg atcgctgcag caccattctc attcatggca aagagcaacc actggatgag
     7201 gaaacaaaag atgcttttca gaatgcctac cttgagttgg gaggccttgg cgagagggtg
     7261 ttaggcttct gtcactttgc tctacctgat gatcagtttc ctgaaggctt ccaatttgat
     7321 gcagatgaag taaactttcc tgtagagaaa ctctgcttta taggactgat gtctatgata
     7381 gaccctcctc gtgctgctgt gccagatgct gttggcaaat gcagaagtgc tggaatcaag
     7441 gttatcatgg ttactggaga ccatccgatc actgccaaag ccattgccaa gggtgttggc
     7501 atcatctctg agggcaatga aacagtagaa gatattgctg ctcggctcaa tattcctgtc
     7561 agccaggtca accctaggga tgcaaaagcc tgtgttgttc atggctcaga tctgaaagac
     7621 atgactagtg agcagttgga tgacatcctg actcatcata cagaaattgt ctttgccagg
     7681 acatctcctc agcagaagct tataattgtg gaaggctgcc agcgacaggg tgctattgta
     7741 gcagtcacag gtgatggtgt gaatgattcc ccagccctga agaaggcaga cattggtgtt
     7801 gcgatgggta ttgctggctc agatgtctcc aagcaggcag ctgacatgat tctgctggat
     7861 gataactttg cctccattgt tactggggtt gaagaaggtc gtctgatctt tgataacctg
     7921 aagaagtcca ttgcttacac cttgaccagt aacattcctg agatcacacc tttcctgatc
     7981 ttcattattg caaacatacc ccttccactg ggaacagtca ctatcctctg cattgatttg
     8041 ggcactgaca tggttcctgc aatctccctg gcatacgagc aagcggagag tgacatcatg
     8101 aagaggcagc ccagaaatcc caaaacagac aagctggtga atgaaagact gatcagcatg
     8161 gcctatgggc agattggtat gatccaggct cttggaggtt tcttcaccta ttttgtaatc
     8221 atggcagaga acgggttctg gccttctggc ttgctaggca tcagagttca atgggatgac
     8281 cgatggatta acgatgtgga agacagctat gggcagcaat ggacttatga acagaggaaa
     8341 attgtggaat tcacttgcca tacagccttc tttgtcagca ttgtggttgt gcaatgggca
     8401 gacttaatca tttgtaagac tcgaaggaac tctgtcttcc agcaggggat gaagaacaag
     8461 attttaatat ttggtctctt tgaggagact gctctggctg ccttcctctc ctactgccct
     8521 gggatggatg ttgctctaag gatgtacccc ctcaaaccga cctggtggtt ctgtgccttc
     8581 ccatactccc tcctcatatt cctgtacgat gaaatcagaa agctgattat cagacgcaac
     8641 cctggtggtt gggtggaaag agagacttac tactaagcgg ccgcgagctc ggagcacaag
     8701 actggcctca tgggccttcc gctcactgcc cgctttccag tgcggccgct ttcgaatcta
     8761 gagcctgcag tctcgacaag cttgtcgaga agtactagag gatcataatc agccatacca
     8821 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     8881 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     8941 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     9001 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
     9061 ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc
     9121 gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat
     9181 ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg
     9241 gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg
     9301 tcatcttcgg ctacttt
//
