LOCUS       sequence-384593-        8330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8330)
  AUTHORS   Chen Z, Zakrzewska S, Hajare HS, Alvarez-Buylla A, Abderemane-Ali F,
            Bogan M, Ramirez D, O'Connell LA, Du Bois J, Minor DL Jr
  TITLE     Definition of a saxitoxin (STX) binding code enables discovery and 
            characterization of the anuran saxiphilin family.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Nov;119(44):e2210114119. doi: 
            10.1073/pnas.2210114119. Epub 2022 Oct 24.
  PUBMED    36279441
REFERENCE   2  (bases 1 to 8330)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-11"; volume: "119(44):e2210114119. 
            doi"; pages: " 10.1073/pnas.2210114119. Epub 2022 Oct 24"
COMMENT     Sequence Label: pFastBac1-RiSxph
FEATURES             Location/Qualifiers
     source          1..8330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        156..247
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     172..189
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3127..3150
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     3816..3837
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3892..3921
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    4072..4206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4109..4128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4163..4182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4235..4400
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4584..5038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4671..4690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4880..4901
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5064..5168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5169..5237,5238..6029)
                     /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(5387..5406)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6200..6788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6689..6708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6942..6959
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  7093..7317
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7384..7917)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(8106..8111,8112..8128,8129..8134))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ggaagacctc ggccgtcgcg gcgcttgccg gtggtgctga ccccggatga agtggttcgc
       61 atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc aggactctag ctatagttct
      121 agtggttggc tacgtatact ccggaatatt aatagatcat ggagataatt aaaatgataa
      181 ccatctcgca aataaataag tattttactg ttttcgtaac agttttgtaa taaaaaaacc
      241 tataaatatt ccggattatt cataccgtcc caccatcggg cgcggatccc ggtccgaagc
      301 gcgcggaatt caaaggccta cgtcgacgag ctcactagtc gcggccgcat gcgaagatcg
      361 tcttacgctg ccagtcaaga aatgagaatg actttgcggg tcctactatg cctgtgtttg
      421 tgggccctca gtagtgccgc cccgaacacg cgaaatgtga ggtggtgtgt cacttcagac
      481 actgaagaac aaaaatgcaa cgaccttgta acttcgtgcc atgttgacga gattctctta
      541 atatgtgtga agaagagtag tacagaagac tgcgtcaggg ccatatcaaa tggcgaagcg
      601 gatgcgatca gtcttgactc gaaagatgtt tataaggcgt ccctggaccc ttttaatctt
      661 aaacccatta tgaccgaggc atactcggag tcagaacata ctccgtgcat gcgacaccgc
      721 caatcagtgc tgggggggaa gatcatgaaa ataggcgctt ttgtcccaaa atgcgatgaa
      781 aagggcaatt atgtacctaa acagtgtcac ggttctactg ggtattgttg gtgcctcaac
      841 gagaacgggg aggaaatcga gggtacgcgt acgcctccag gaaatccagg gttgacatgt
      901 gaaaataaag ctaataagcc cccttgtctc aaagagcgcc aaaaactcct tagcgccaaa
      961 ccatccccag ctgtatttgt ccctgaatgt gacgagaagg gtaactatcg accacagcag
     1021 agccatgtct atagctggtg tgttgatgag tatggtgaag aagtattcgg cagtaggagt
     1081 ttccctggta agcctcctaa accctgcgag gcttcaggcg aaaccccctg tataaaagaa
     1141 aggaataaag tactttcggc ggcagagcct ttgctgggtg cctttgtacc tgattgcgat
     1201 gagaagggat atttctctcc actacagtca cacgggtcta ccggatatag ctggtgtgta
     1261 actaagaacg gggatgagat taagggcacc cgcacggggc ccggacagtc cccaccaaca
     1321 tgtgaggtgc ccgcgccagt gacacttcat tatgctgtag cagttgttaa aaaatcttcg
     1381 agctttcacc tagaccaact taagggtaaa agatcatgtc acagtgctgt cggcgaagcg
     1441 gccgggtggg ttgctcccct taacgtcctt ttgaagaaga agctcctact gtgggaggaa
     1501 ccggagccga agaccataga aaaagtggca tcagagattt tttctgccag ttgtgccccc
     1561 ggcgcgcaag aggcgaattt atgtgagcag tgtgcaggtc aagaggacaa atgcaccaga
     1621 gggccgggtg agccgtacta cggagacgaa ggtgcctttc gatgcttaag ggaggacaaa
     1681 ggtgacgtcg cgtttgtgga agacactgct ctaaccggcc aatacagtga taacttcgaa
     1741 ttgttatgcc cggataatac tcgcaggccc ttatctcaat acaaaatatg caatttcggt
     1801 cgcataccgc gacacgcagt tgttacgcgg tcttccggag acaaaatgaa agacataaca
     1861 gagtatttac tacaagcgca aaagaaggaa tgcaaactgt tctcgtctac ccacggaaag
     1921 gacctattat ttgaggacac gacgagcgca ctcatagcac tgccgtctgc gatggatacc
     1981 tttttgtttt taggacctga tctgtttaac ggcatgaaaa cactcaacgg agcacgtccg
     2041 ccccaggtaa accagcagat cagatggtgc ccacagagca aaaacgagaa gaagaaatgc
     2101 gatgattgga gcagcgtttc cgggggggca attaaatgca cggaaccgtc gtctggactg
     2161 gaatgcatca agatgatcct caagggcgag gctgaagctg ttaacttaga tatagagcat
     2221 tcatatatcg cactgaaatg tggactctta ccatcagttg acgagtatcg gaataaggat
     2281 gatcttatcc catgccaaat cccgggcagt gaatacagcg aattcggttc ttaccggatt
     2341 gttgccctcg taaaaaagac cgataaggat ctaacatgga acaatctgca agggaagaag
     2401 tcgtgccata cccgcgcagg cgatatgatc ggttggaata ttcccgtgta cttaatatca
     2461 aagaagacaa aaaactgtga tcttgggagc tacttcagcc aatcgtgtgc gccaggttct
     2521 cctatcgaca gtaacctgtg taaactttgt ataggcgacc cccaaaatcc tcaagccaat
     2581 acaaagtgca gcatttctga caaggaagcg tactacggac acgagggagc catacgctgt
     2641 ctagtggaaa aaggcgacgt ggctttcgtg cctcatacag caattttcga gaacacagac
     2701 ggtaagaacc cggccctctg ggcaaaggac ctaaaaagta ctgatttcga gttattgtgc
     2761 ccggacggat cacgggctcc cgtgaatgac tacaaaaagt gcaagttgtc cgggattgcg
     2821 aaccagattg caatcacgcg gccggaaagt gtcaaggacg tagtacgtat tatacaaaat
     2881 cagcagtccc tgtacggtcg tacggggtcc cagaaagata ttttccagat gttctcttcg
     2941 tcctatggac agaatctctt gttctccgat cgtactcagt gccttgtcga atttgacaga
     3001 atgctagaca gggatatcat ggatgattat tttggcaaac ctttccataa acacctccta
     3061 cgtgataatg attgcttgcc gaagtccgcg ttggcgtccg cttgtagttt tcaccatagc
     3121 aactctctgg aggtcctgtt ccaaggacca actgctgccg ctgccgtgtc taagggcgaa
     3181 gagctgttca ctggagtggt ccctatcctg gtcgagctgg acggtgacgt gaacggccac
     3241 aagttctcag tctccggtga aggcgaggga gacgctacct acggaaagct gactctgaag
     3301 ttcatctgca ccactggcaa gctgccagtc ccttggccca ccctggtgac cactctgact
     3361 tacggtgtgc agtgcttcag ccgctaccct gaccacatga agcagcacga cttcttcaag
     3421 tctgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggaaac
     3481 tacaagaccc gtgctgaagt caagttcgag ggcgacactc tggtgaacag gatcgaactg
     3541 aagggaatcg acttcaagga ggacggaaac atcctgggtc acaagctgga atacaactac
     3601 aactcacaca acgtctacat catggccgac aagcagaaga acggtatcaa ggtgaacttc
     3661 aagatcagac acaacatcga ggacggctcc gtccagctgg ctgaccacta ccagcagaac
     3721 actcctatcg gtgacggacc tgtgctgctg cccgacaacc actacctgtc cactcagagc
     3781 aagctgtcta aggacccaaa cgaaaagcgc gaccacatgg tcctgctgga gttcgtgacc
     3841 gctgccggca tcactctggg aatggacgaa ctctacaaat ctggcggcgg acatcatcac
     3901 caccaccatc atcaccacca ctaactcgag gcatgcggta ccaagcttgt cgagaagtac
     3961 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4021 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4081 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4141 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4201 tggatctgat cactgcttga gcctaggaga tccgaaccag ataagtgaaa tctagttcca
     4261 aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct
     4321 attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc
     4381 aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat
     4441 cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga
     4501 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat
     4561 ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4621 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4681 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4741 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4801 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4861 ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4921 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4981 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     5041 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5101 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5161 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5221 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5281 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5341 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5401 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5461 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5521 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5581 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5641 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5701 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5761 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5821 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5881 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5941 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6001 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6061 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6121 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6181 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6241 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6301 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6361 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6421 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6481 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6541 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6601 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6661 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6721 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6781 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6841 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6901 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6961 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     7021 gcatagacca gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc
     7081 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
     7141 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     7201 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     7261 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg
     7321 gcgttgtgac aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa
     7381 ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa
     7441 ctttgtatag agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata
     7501 agcaccaagc gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg
     7561 cctccggtgc tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc
     7621 aaacttgggc agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc
     7681 aagcgcgatg aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg
     7741 ttgggagtag gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat
     7801 ggatttgact tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc
     7861 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt
     7921 tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc
     7981 gaggcataga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt
     8041 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc
     8101 attacagttt acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc
     8161 acggtgtgcg tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc
     8221 tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt
     8281 tcttctacgg caaggtgctg tgcacggatc tgccctggct tcaggagatc
//
