LOCUS       sequence-394224-        8194 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 8194)
  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 8194)
  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-OsSxph
FEATURES             Location/Qualifiers
     source          1..8194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        172..263
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     188..205
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3041..3064
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3080..3082,3083..3793)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3383..3402)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3730..3751
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3806..3835
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    3971..4105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4008..4027)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4062..4081
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4134..4299
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4483..4937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4570..4589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4779..4800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4963..5067
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5068..5136,5137..5928)
                     /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(5286..5305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6099..6687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6588..6607
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6841..6858
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6992..7216
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7283..7816)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7986..7991,7992..8008,8009..8014))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc
       61 ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga
      121 ctctagctat agttctagtg gttggctacg tatactccgg aatattaata gatcatggag
      181 ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt
      241 ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc atcggatgcg
      301 tatgacgctg caagtattgt tgtgcctttg tctcttggta ctcagttcag cagcgcctaa
      361 ttcaagaaat gtgagatggt gcgtcaccag tgataccgag gagcagaaat gtaatgatct
      421 ggtcacttct tgtcatgtgg atgaaatcct gctcatctgt gttaaaaaat cttcgaagga
      481 agattgcgta cgtgcgatct ctaacggtga agctgatgcg atatctctgg actcgaaaga
      541 tgtctacaag gctagcttgg atccgtttaa cctcaaaccg atcatgaccg aagcctattc
      601 tgaatccgaa cagaccccat gcatgcgtca tagacaatct gttttgggcg gtaaaaaaat
      661 gattattggc gcatttgttc cgaaatgcga cgaaaaaggt aactatgtcc ccaaacaatg
      721 tcacgactca actggctttt gctggtgtct caatgaaaac ggcgaagaga ttgagggaac
      781 taggactccc ccgggaaatt caggtcttac ctgtgaaaat aaggcgaata agcccccttg
      841 tctcaaagag cgtcagaagc tcctctctgc aaaacccagt cccgcagtat ttgttcccga
      901 atgcgatgag aagggtaact atcgcccaga gcagtcccat gtgtattcct ggtgcgtcga
      961 tgagtacgga gaggaggttc ttggctcacg tacgttccct ggtaagcccc ccaagccatg
     1021 cgaggcttcg ggtgaaacgc cctgtattaa agagcgcaac aaggttctca gcgccgctga
     1081 acccctgcca ggcgcctttg taccagattg tgatgaaaaa ggttatttct cccccctcca
     1141 gttccacgga tcgacgggcc acagttggtg tgtgaccaaa aatggagagg aaattaaagg
     1201 tacgaggaca aggccgggtc aaacgccgcc aacctgtgat gtaccagccc ctgcaacatt
     1261 gcactacgct gtagccgttg tgaaaaagag ctctagtttt cagctcgacc agcttaaggg
     1321 aaaacgctcg tgtcatagcg ctgtaggtga ggctgcggga tgggtagcac cacttaatgt
     1381 gcttctgaaa aaaaagtttc tcttgtggga ggaacctgaa caaaaatcga ctgagaaggt
     1441 ggcatccgag atattcagtg cgtcttgcgc agcaggcgcc caagaagcga atctctgtga
     1501 gcaatgcgcc ggacaagaag acaagtgcac ccgtggtcca ggcgaaccgt attatggtga
     1561 cgaaggagcc ataagatgcc ttagagatga taaaggtgat gtagctttcg tagaggacac
     1621 tgcattgacc ggccaatact cggataactt cgagctcctg tgccccgaca acacacgccg
     1681 tcctctctca cagtacaaga tatgtaattt tggttccgtg ccgaggcacg cggtagtcac
     1741 cagatccaca ggcgataaaa tgaaggaaat aacggagtac ctgctccagg cgcaaaagaa
     1801 agaatgcaag ttgttcagca gcacccacgg taaggacctc ctgtttgaag atacgacgtc
     1861 tgcgttgatc gcacttccgt cgggaatgga tactttcttg tttttgggcc ctgatttgtt
     1921 caacggaatg aagacattga atggtgccca ccctcctagg gtaaaacaga aaattaggtg
     1981 gtgcccccaa aataaaaacg aaaaaaaaaa gtgtgacgac tggtcgtccg tttcgggagg
     2041 ttccataaag tgcacagagc cttcaagcgc attggaatgc attaagatga tcctcaagga
     2101 tgaagccgat gctgttaact tggatgcgga gcacgcgtat acagctctca aatgcggatt
     2161 gcttccttct cttgaagagt ataggaacaa agatgacttg attgtctgtc agatcccagg
     2221 cgcagaatat agtgagtttg gatcatacag gatcgttgcc cttgttaaaa aaacagataa
     2281 ggacataact tggaataatc tccagggtaa gaaaagttgt catacccacg tcggcgatat
     2341 gatcggttgg aacatacctg tgtatttgat aagtaaaaag actaagaatt gcgatctggg
     2401 ctcatatttt tcccaatctt gtgcaccggg cagccccata gactcgaacc tctgcaagct
     2461 ctgtattggc gacccacaga acactgaggc taacacaaag tgctcgttgt cagacaagga
     2521 ggcttactat ggcaatgagg gtgccgttcg ttgccttgtc gaaacaggtg acgtcgcctt
     2581 tgtgccccac acagctgttt ttgagaacac tgacggaaaa aatcctgctt tgtgggcaaa
     2641 ggaccttaaa tctacggact tcgaattgct ttgccccgac ggctctagag ctcctgtgtc
     2701 ggactataag aagtgtaagc tgtcaggcat tgccaaccat gtaacggtga caaggccgga
     2761 aagcgtgaaa gacgtagtga ggatcactca gaaccaggag agtctgtacg gaagaacggg
     2821 ttcccaaaag gacattttcc aaatgtttag ctccagctac ggtcaaaacc ttctgttcaa
     2881 cgataggacc cagtgtcttt ttgaatttga ccgcatgctg gatagggata taatggatga
     2941 ttatttgggc aagccctttc acaaacacct tatgtcagac aacgattgcc tccctaaaag
     3001 tgccttggcc actgcgtgca gctttcacca ctcgaattct ttggaagttt tgtttcaagg
     3061 tccaactgct gccgccgctg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3121 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3181 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3241 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3301 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3361 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3421 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3481 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3541 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3601 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3661 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
     3721 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3781 cgagctgtac aagtccggag gtggtcacca tcaccatcac caccatcacc atcactaata
     3841 gaagcttgtc gagaagtact agaggatcat aatcagccat accacatttg tagaggtttt
     3901 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3961 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4021 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4081 caatgtatct tatcatgtct ggatctgatc actgcttgag cctaggagat ccgaaccaga
     4141 taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg
     4201 ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt
     4261 tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt
     4321 tatgttttta atcaaacatc ctgccaactc catgtgacaa accgtcatct tcggctactt
     4381 tttctctgtc acagaatgaa aatttttctg tcatctcttc gttattaatg tttgtaattg
     4441 actgaatatc aacgcttatt tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     4501 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4561 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4621 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4681 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4741 tttttcgccc tttgacgttg gagtccacgt tcttaatagt ggactcttgt tccaaactgg
     4801 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4861 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4921 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4981 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5041 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5101 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5161 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5221 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5281 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5341 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5401 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5461 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5521 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5581 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5641 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5701 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5761 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5821 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5881 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5941 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6001 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6061 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6121 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6181 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6241 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6301 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6361 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6421 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6481 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6541 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6601 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6661 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6721 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6781 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6841 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     6901 ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt
     6961 gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga
     7021 acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg
     7081 aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac
     7141 tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat
     7201 ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc
     7261 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     7321 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     7381 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     7441 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     7501 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     7561 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     7621 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     7681 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     7741 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     7801 tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc
     7861 gaggcataga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt
     7921 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc
     7981 attacagttt acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc
     8041 acggtgtgcg tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc
     8101 tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt
     8161 tcttctacgg caaggtgctg tgcacggatc tgcc
//
