LOCUS       sequence-384668-        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-NpSxphI559Y
FEATURES             Location/Qualifiers
     source          1..8096
                     /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             2893..2916
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     3582..3603
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3658..3687
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    3838..3972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3875..3894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3929..3948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4001..4166
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4350..4804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4437..4456)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4646..4667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4830..4934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4935..5003,5004..5795)
                     /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(5153..5172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5966..6554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6455..6474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6708..6725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6859..7083
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7150..7683)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7872..7877,7878..7894,7895..7900))
                     /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 ggcgttgacc
      361 ttccatacag ccctctactt cacgattgtt ggtttgagtt tcgcagcatc tgacgctagg
      421 catgtccagt ggtgtaccat cagccacctg gaacagaaga agtgcaacga cctggtcggc
      481 tcttgcaacg tgccagacat caccctggct tgcgtgtacc gttccagcac tgagaactgc
      541 atggctgcca tcaaggacgg acaggctgac gccatgttcc tggacagcgg tgacgtctac
      601 aaggcctctc tggaccacta caacctgaag ccaatcatcg ctgaacctta ctcactgcac
      661 agggagctga ctaagtgcct gaagcacaga caggaatccc tgggtggcga caagatggtg
      721 aagggcaggt acatcccaca gtgcgacgag aagggaaact accaccctgt ccagtgccac
      781 gcctcaaccg gctactgctg gtgcgtgaac gctaacggtg aaaagatcga gggcactaac
      841 accactcctg tccagacccc tcccacttgc ccctcccagg tgctgaccaa gtgcctgaag
      901 gaaagacagg aggccctggg tggcaagcgc atcgctatcg gtcgttacat cccccagtgc
      961 gacgagcagg gcaactaccg cccaatgcag tgccacggaa gcactggtta ctgttggtgt
     1021 gtgaacgcca tcggagaaaa gatcgagggt accaacactc cacctggtaa cacccagcct
     1081 acttgccagt ctcacgactg ggacacctgc cactacgctg tcgccgtggt caagaactct
     1141 tcaactttcc agttcggcca gctgaaggga aagcgtagct gccactctgg tctgtcaaag
     1201 accgacggct ggaacgctcc tgtgaacgtc ttcgtggaaa agaagctcct cccatgggac
     1261 ggtctggcta agggatcaat cgagagggct gtgtccaagt tcttctccgc cagctgcatc
     1321 cctggtgcta ccgagactaa cctgtgcaag cagtgcatcg gcgaggaaga gaagaagtgc
     1381 aagtccagcc acgacgaacc ctactacggc gaccacggag ctttccgttg cctgcaggag
     1441 gacaagggag acgtggcctt cctgaagaac accgctctgc cagacgaaca ctccggtgtc
     1501 tacgagctgc tgtgcccaga caacactcgt aagcctctga acaagtacaa ggaatgcaac
     1561 ctgggaaagg tgcctgctga cgccgtggtc accaggaagg ccggtgacaa gactaaggac
     1621 atcaacgact tcctgctgga ggctcagaag aagaagtgca agctgttcgg aagccctcac
     1681 ggcaaggacc tgatgttcga cgactcaacc actcacctgg cccctctgcc ctccgaaatc
     1741 gacgctttct tcttcctggg tgtcaagtgg tacaacgcta tgaaggccct gaccgaagac
     1801 gtcaagctgc catctaagaa caaggtgaga tggtgcacta tcaacaagcc tgagatgatg
     1861 aagtgcaagg actgggctgc cgtgagcggc ggagctatcg cttgcaccga agcctcttgc
     1921 cccgagcact gcgtcaagca gatcctgaag ggagaggctg acgccgtcac tctggacgtg
     1981 cagtacatgt acatggctct gatgtgcggc ctgctgccag ctgtggaaga gtacccaaac
     2041 aaggacgact tccacccctg ccagatccca ggtagcacct acaaggactt cggcactaag
     2101 cgtgctgtcg ccctggtgaa gaagtcaaac aaggacatca agtggaacaa cctgaaggga
     2161 aagaagtcct gccacaccca cgtgggcgac atcccaggat gggtcatccc agccggtctg
     2221 atctcaaacc agaacgacaa catcgacatc gaaagcttct tcggagagtc ttgcgctccc
     2281 ggttcagaca ccaactccaa gctgtgcaag ctgtgcatcg gtgacccaga gaaccctaag
     2341 gcttccacta gatgctctct gtcagacaag gaagcctact acggtaacga gggcgctttc
     2401 cgctgcctgg tggaaaaggg cgacgtggct ttcgtgccac acaccgtggt cttcgctaac
     2461 actgacggaa agaacccagc tgagtgggcc aaggacctga agtcagaaga cttcgagatc
     2521 ctctgcctgg acggctcccg cgccccagtc accaactaca ggggatgcaa cctgtctgga
     2581 ctgcccccaa gggctatcgt cactagagaa gagtccgtga gcgacgtggt caggatcctg
     2641 atcaaccagc agtctctgta cggaagaaac ggtttcgaaa aggacatgtt ccagatgttc
     2701 tcttcagcca agggccagaa cctgctgttc aacgacgaaa cccagtgcct gatcgagttc
     2761 gacaggcagc caaaggacat catggaggac tacttcggag tccgctacta caccgctgtg
     2821 tactctgcct cacgttccgc tgtgcctagc gaactgatcc ccgcttgcac tttcaagcac
     2881 tgcagcaact ctctggaggt cctgttccaa ggaccaactg ctgccgctgc cgtgtctaag
     2941 ggcgaagagc tgttcactgg agtggtccct atcctggtcg agctggacgg tgacgtgaac
     3001 ggccacaagt tctcagtctc cggtgaaggc gagggagacg ctacctacgg aaagctgact
     3061 ctgaagttca tctgcaccac tggcaagctg ccagtccctt ggcccaccct ggtgaccact
     3121 ctgacttacg gtgtgcagtg cttcagccgc taccctgacc acatgaagca gcacgacttc
     3181 ttcaagtctg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3241 ggaaactaca agacccgtgc tgaagtcaag ttcgagggcg acactctggt gaacaggatc
     3301 gaactgaagg gaatcgactt caaggaggac ggaaacatcc tgggtcacaa gctggaatac
     3361 aactacaact cacacaacgt ctacatcatg gccgacaagc agaagaacgg tatcaaggtg
     3421 aacttcaaga tcagacacaa catcgaggac ggctccgtcc agctggctga ccactaccag
     3481 cagaacactc ctatcggtga cggacctgtg ctgctgcccg acaaccacta cctgtccact
     3541 cagagcaagc tgtctaagga cccaaacgaa aagcgcgacc acatggtcct gctggagttc
     3601 gtgaccgctg ccggcatcac tctgggaatg gacgaactct acaaatctgg cggcggacat
     3661 catcaccacc accatcatca ccaccactaa ctcgaggcat gcggtaccaa gcttgtcgag
     3721 aagtactaga ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3781 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3841 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3901 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3961 catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa gtgaaatcta
     4021 gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta cacccagttc
     4081 ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc acccctccca
     4141 gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat gtttttaatc
     4201 aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt ctctgtcaca
     4261 gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact gaatatcaac
     4321 gcttatttgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     4381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4501 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4561 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4621 gacgttggag tccacgttct taatagtgga ctcttgttcc aaactggaac aacactcaac
     4681 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4741 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4801 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4861 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4921 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4981 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5041 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5101 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5161 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5221 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5281 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5341 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5401 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5461 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5521 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5581 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5641 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5701 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5761 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5821 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5881 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5941 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6001 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6061 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6121 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6181 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6241 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6301 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6361 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6421 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6481 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6541 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6601 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6661 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6721 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     6781 cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag taataaatgg
     6841 atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca aaatagatct
     6901 aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa tcagtccagt
     6961 tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct tcattttctg
     7021 aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt aaagactata
     7081 ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg
     7141 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
     7201 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
     7261 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
     7321 gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg
     7381 ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     7441 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     7501 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     7561 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     7621 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa
     7681 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     7741 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     7801 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     7861 acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc
     7921 gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt
     7981 cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct
     8041 tgctgttctt ctacggcaag gtgctgtgca cggatctgcc ctggcttcag gagatc
//
