LOCUS       sequence-162901-        6354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus Expression of LFn-Needle fusion. The LFn domain acts as 
            a signal sequence that targets proteins for cytosolic translocation 
            via the co-administered protective antigen (PA) channel protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6354)
  AUTHORS   Rauch I, Tenthorey JL, Nichols RD, Al Moussawi K, Kang JJ, Kang C, 
            Kazmierczak BI, Vance RE
  TITLE     NAIP proteins are required for cytosolic detection of specific 
            bacterial ligands in vivo.
  JOURNAL   J Exp Med. 2016 May 2;213(5):657-65. doi: 10.1084/jem.20151809. Epub
            2016 Apr 4.
  PUBMED    27045008
REFERENCE   2  (bases 1 to 6354)
  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"; doi: 
            "10.1084/jem.20151809"; journalName: "J Exp Med"; date: "2016-05-2- 
            2"; volume: "213"; issue: "5"; pages: "657-65"
COMMENT     Sequence Label: pFastBacDual with LFn + Needle
FEATURES             Location/Qualifiers
     source          1..6354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..557
                     /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     399..420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        584..688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(689..757,758..1549)
                     /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(907..926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1720..2308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2209..2228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2462..2479
                     /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
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     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    4161..4209
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4234..4253
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4345..4454)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4478..4569
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4494..4511
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4663..4680
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4690..4707
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     polyA_signal    5944..6078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5981..6000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6035..6054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6107..6272
                     /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 ctttaatagt ggactcttgt tccaaactgg
      421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      541 attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2101 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2461 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2521 tctgtgcggt atttcacacc gcagaccagc 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 acctgggcag 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 ccctggcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcct acgtacccgt agtggctatg gcagggcttg ccgccccgac
     4021 gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga
     4081 aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg
     4141 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt
     4201 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     4261 tcccccatct cccggtaccg catgctatgc atcagctgct agcaccatgg ctcgagatcc
     4321 cgggtgatca agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt
     4381 taattaatat acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg
     4441 ttgaattaaa ggtccgtata ctccggaata ttaatagatc atggagataa ttaaaatgat
     4501 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa
     4561 cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc ccggtccgaa
     4621 gcgcgcggaa ttcaaaggcc tacgtcgaca atgggcagca gccatcatca tcatcatcac
     4681 agcagcggcc tggtgccgcg cggcagccat atggcgggcg gtcatggtga tgtaggtatg
     4741 cacgtaaaag agaaagagaa aaataaagat gagaataaga gaaaagatga agaacgaaat
     4801 aaaacacagg aagagcattt aaaggaaatc atgaaacaca ttgtaaaaat agaagtaaaa
     4861 ggggaggaag ctgttaaaaa agaggcagca gaaaagctac ttgagaaagt accatctgat
     4921 gttttagaga tgtataaagc aattggagga aagatatata ttgtggatgg tgatattaca
     4981 aaacatatat ctttagaagc attatctgaa gataagaaaa aaataaaaga catttatggg
     5041 aaagatgctt tattacatga acattatgta tatgcaaaag aaggatatga acccgtactt
     5101 gtaatccaat cttcggaaga ttatgtagaa aatactgaaa aggcactgaa cgtttattat
     5161 gaaataggta agatattatc aagggatatt ttaagtaaaa ttaatcaacc atatcagaaa
     5221 tttttagatg tattaaatac cattaaaaat gcatctgatt cagatggaca agatctttta
     5281 tttactaatc agcttaagga acatcccaca gacttttctg tagaattctt ggaacaaaat
     5341 agcaatgagg tacaagaagt atttgcgaaa gcttttgcat attatatcga gccacagcat
     5401 cgtgatgttt tacagcttta tgcaccggaa gcttttaatt acatggataa atttaacgaa
     5461 caagaaataa atctatcctt ggaagaactt aaagatcaac ggagcacgcg tagcaatccc
     5521 ccgactccat tgctgaccga ctacgagtgg tcaggctacc tgaccggcat cggccgcgcg
     5581 ttcgacaccg gcgtgaagga tctgaaccag caactgcagg acgcgcaggc gaatctcacg
     5641 aagaacccga gcgatccgac ggcgctcgcg aactatcaga tgatcatgtc ggaatacaac
     5701 ctgtaccgga acgcgcagag cagcgcggtc aaatcgatga aggacatcga ttcgtcgatc
     5761 gtctcgaact tccgctaagc ggccgctttc gaatctagag cctgcagtct cgacaagctt
     5821 gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     5881 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     5941 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6001 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6061 tcttatcatg tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga
     6121 aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc
     6181 cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc
     6241 ctcccagttc ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt
     6301 ttaatcaaac atcctgccaa ctccatgtga caaaccgtca tcttcggcta cttt
//
