LOCUS       sequence-394165-        8096 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 8096)
  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 8096)
  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-NpSxph
FEATURES             Location/Qualifiers
     source          1..8096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        178..269
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     194..211
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2915..2938
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     3604..3625
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3680..3709
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    3860..3994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3897..3916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3951..3970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4023..4188
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4372..4826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4459..4478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4668..4689
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4852..4956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4957..5025,5026..5817)
                     /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(5175..5194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5988..6576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6477..6496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6730..6747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6881..7105
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7172..7705)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7894..7899,7900..7916,7917..7922))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tctgccctgg cttcaggaga tcggaagacc tcggccgtcg cggcgcttgc cggtggtgct
       61 gaccccggat gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc
      121 ccaggactct agctatagtt ctagtggttg gctacgtata ctccggaata ttaatagatc
      181 atggagataa ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta
      241 acagttttgt aataaaaaaa cctataaata ttccggatta ttcataccgt cccaccatcg
      301 ggcgcggatc ccggtccgaa gcgcgcggaa ttcaaaggcc tacgtcgacg agctcactag
      361 tcgcggccgc atggcgttga ccttccatac agccctctac ttcacgattg ttggtttgag
      421 tttcgcagca tctgacgcta ggcatgtcca gtggtgtacc atcagccacc tggaacagaa
      481 gaagtgcaac gacctggtcg gctcttgcaa cgtgccagac atcaccctgg cttgcgtgta
      541 ccgttccagc actgagaact gcatggctgc catcaaggac ggacaggctg acgccatgtt
      601 cctggacagc ggtgacgtct acaaggcctc tctggaccac tacaacctga agccaatcat
      661 cgctgaacct tactcactgc acagggagct gactaagtgc ctgaagcaca gacaggaatc
      721 cctgggtggc gacaagatgg tgaagggcag gtacatccca cagtgcgacg agaagggaaa
      781 ctaccaccct gtccagtgcc acgcctcaac cggctactgc tggtgcgtga acgctaacgg
      841 tgaaaagatc gagggcacta acaccactcc tgtccagacc cctcccactt gcccctccca
      901 ggtgctgacc aagtgcctga aggaaagaca ggaggccctg ggtggcaagc gcatcgctat
      961 cggtcgttac atcccccagt gcgacgagca gggcaactac cgcccaatgc agtgccacgg
     1021 aagcactggt tactgttggt gtgtgaacgc catcggagaa aagatcgagg gtaccaacac
     1081 tccacctggt aacacccagc ctacttgcca gtctcacgac tgggacacct gccactacgc
     1141 tgtcgccgtg gtcaagaact cttcaacttt ccagttcggc cagctgaagg gaaagcgtag
     1201 ctgccactct ggtctgtcaa agaccgacgg ctggaacgct cctgtgaacg tcttcgtgga
     1261 aaagaagctc ctcccatggg acggtctggc taagggatca atcgagaggg ctgtgtccaa
     1321 gttcttctcc gccagctgca tccctggtgc taccgagact aacctgtgca agcagtgcat
     1381 cggcgaggaa gagaagaagt gcaagtccag ccacgacgaa ccctactacg gcgaccacgg
     1441 agctttccgt tgcctgcagg aggacaaggg agacgtggcc ttcctgaaga acaccgctct
     1501 gccagacgaa cactccggtg tctacgagct gctgtgccca gacaacactc gtaagcctct
     1561 gaacaagtac aaggaatgca acctgggaaa ggtgcctgct gacgccgtgg tcaccaggaa
     1621 ggccggtgac aagactaagg acatcaacga cttcctgctg gaggctcaga agaagaagtg
     1681 caagctgttc ggaagccctc acggcaagga cctgatgttc gacgactcaa ccactcacct
     1741 ggcccctctg ccctccgaaa tcgacgcttt cttcttcctg ggtgtcaagt ggtacaacgc
     1801 tatgaaggcc ctgaccgaag acgtcaagct gccatctaag aacaaggtga gatggtgcac
     1861 tatcaacaag cctgagatga tgaagtgcaa ggactgggct gccgtgagcg gcggagctat
     1921 cgcttgcacc gaagcctctt gccccgagca ctgcgtcaag cagatcctga agggagaggc
     1981 tgacgccgtc actctggacg tgcagtacat gtacatggct ctgatgtgcg gcctgctgcc
     2041 agctgtggaa gagtacccaa acaaggacga cttccacccc tgccagatcc caggtagcac
     2101 catcaaggac ttcggcacta agcgtgctgt cgccctggtg aagaagtcaa acaaggacat
     2161 caagtggaac aacctgaagg gaaagaagtc ctgccacacc cacgtgggcg acatcccagg
     2221 atgggtcatc ccagccggtc tgatctcaaa ccagaacgac aacatcgaca tcgaaagctt
     2281 cttcggagag tcttgcgctc ccggttcaga caccaactcc aagctgtgca agctgtgcat
     2341 cggtgaccca gagaacccta aggcttccac tagatgctct ctgtcagaca aggaagccta
     2401 ctacggtaac gagggcgctt tccgctgcct ggtggaaaag ggcgacgtgg ctttcgtgcc
     2461 acacaccgtg gtcttcgcta acactgacgg aaagaaccca gctgagtggg ccaaggacct
     2521 gaagtcagaa gacttcgaga tcctctgcct ggacggctcc cgcgccccag tcaccaacta
     2581 caggggatgc aacctgtctg gactgccccc aagggctatc gtcactagag aagagtccgt
     2641 gagcgacgtg gtcaggatcc tgatcaacca gcagtctctg tacggaagaa acggtttcga
     2701 aaaggacatg ttccagatgt tctcttcagc caagggccag aacctgctgt tcaacgacga
     2761 aacccagtgc ctgatcgagt tcgacaggca gccaaaggac atcatggagg actacttcgg
     2821 agtccgctac tacaccgctg tgtactctgc ctcacgttcc gctgtgccta gcgaactgat
     2881 ccccgcttgc actttcaagc actgcagcaa ctctctggag gtcctgttcc aaggaccaac
     2941 tgctgccgct gccgtgtcta agggcgaaga gctgttcact ggagtggtcc ctatcctggt
     3001 cgagctggac ggtgacgtga acggccacaa gttctcagtc tccggtgaag gcgagggaga
     3061 cgctacctac ggaaagctga ctctgaagtt catctgcacc actggcaagc tgccagtccc
     3121 ttggcccacc ctggtgacca ctctgactta cggtgtgcag tgcttcagcc gctaccctga
     3181 ccacatgaag cagcacgact tcttcaagtc tgccatgccc gaaggctacg tccaggagcg
     3241 caccatcttc ttcaaggacg acggaaacta caagacccgt gctgaagtca agttcgaggg
     3301 cgacactctg gtgaacagga tcgaactgaa gggaatcgac ttcaaggagg acggaaacat
     3361 cctgggtcac aagctggaat acaactacaa ctcacacaac gtctacatca tggccgacaa
     3421 gcagaagaac ggtatcaagg tgaacttcaa gatcagacac aacatcgagg acggctccgt
     3481 ccagctggct gaccactacc agcagaacac tcctatcggt gacggacctg tgctgctgcc
     3541 cgacaaccac tacctgtcca ctcagagcaa gctgtctaag gacccaaacg aaaagcgcga
     3601 ccacatggtc ctgctggagt tcgtgaccgc tgccggcatc actctgggaa tggacgaact
     3661 ctacaaatct ggcggcggac atcatcacca ccaccatcat caccaccact aactcgaggc
     3721 atgcggtacc aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt
     3781 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3841 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3901 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3961 caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc ctaggagatc
     4021 cgaaccagat aagtgaaatc tagttccaaa ctattttgtc atttttaatt ttcgtattag
     4081 cttacgacgc tacacccagt tcccatctat tttgtcactc ttccctaaat aatccttaaa
     4141 aactccattt ccacccctcc cagttcccaa ctattttgtc cgcccacagc ggggcatttt
     4201 tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt
     4261 cggctacttt ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt
     4321 ttgtaattga ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct
     4381 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4441 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4501 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4561 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4621 gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt
     4681 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4741 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4801 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     4861 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4921 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4981 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5041 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5101 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5161 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5221 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5281 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5341 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5401 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5461 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5521 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5581 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5641 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5701 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5761 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5821 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5881 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5941 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6001 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6061 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6121 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6181 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6241 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6301 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6361 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6421 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6481 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6541 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6601 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6661 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6721 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     6781 cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg
     6841 gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc
     6901 ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt
     6961 tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct
     7021 aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc
     7081 caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg
     7141 ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa
     7201 agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac
     7261 cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga
     7321 gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat
     7381 agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc
     7441 caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt
     7501 tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac
     7561 gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg
     7621 tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg
     7681 taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg
     7741 gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa
     7801 caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac
     7861 cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc
     7921 aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc
     7981 agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg
     8041 catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg cacgga
//
