LOCUS       sequence-309416-        3624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector with FLAG tag expressed under the IE1 promoter and hr5 
            enhancer from Autographa californica nuclear polyhedris virus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3624)
  AUTHORS   Russell TA, Ayaz A, Davidson AD, Fernandez-Sesma A, Maringer K
  TITLE     Imd pathway-specific immune assays reveal NF-kappaB stimulation by 
            viral RNA PAMPs in Aedes aegypti Aag2 cells.
  JOURNAL   PLoS Negl Trop Dis. 2021 Feb 16;15(2):e0008524. doi: 
            10.1371/journal.pntd.0008524.
  PUBMED    33591970
REFERENCE   2  (bases 1 to 3624)
  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: "PLoS Negl 
            Trop Dis."; date: "2021-02-16"; volume: "15(2):e0008524. doi"; 
            pages: " 10.1371/journal.pntd.0008524"
COMMENT     Sequence Label: pKM3
FEATURES             Location/Qualifiers
     source          1..3624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..483
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        487..1078
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     CDS             1087..1110
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1141..1448
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     promoter        1478..1582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1583..1651,1652..2443)
                     /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(1801..1820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2614..3202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3103..3122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3356..3373
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3490..3511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3526..3531,3532..3549,3550..3556)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3564..3580
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3569..3591
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3588..3604
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3588..3604
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgcgtaaaac acaatcaagt atgagtcata agctgatgtc atgttttgca cacggctcat
       61 aaccgaactg gctttacgag tagaattcta cttgtaacgc acgatcagtg gatgatgtca
      121 tttgtttttc aaatcgagat gatgtcatgt tttgcacacg gctcataaac tcgctttacg
      181 agtagaattc tacgtgtaac gcacgatcga ttgatgagtc atttgttttg caatatgata
      241 tcatacaata tgactcattt gtttttcaaa accgaacttg atttacgggt agaattctac
      301 ttgtaaagca caatcaaaaa gatgatgtca tttgtttttc aaaactgaac tcgctttacg
      361 agtagaattc tacgtgtaaa acacaatcaa gaaatgatgt catttgttat aaaaataaaa
      421 gctgatgtca tgttttgcac atggctcata actaaactcg ctttacgggt agaattctac
      481 gcgcgtcgat gtctttgtga tgcgcgcgac atttttgtag gttattgata aaatgaacgg
      541 atacgttgcc cgacattatc attaaatcct tggcgtagaa tttgtcgggt ccattgtccg
      601 tgtgcgctag catgcccgta acggacctcg tacttttggc ttcaaaggtt ttgcgcacag
      661 acaaaatgtg ccatacttgc agctctgcat gtgtgcgcgt taccacaaat cccaacggcg
      721 cagtgtactt gttgtatgca aataaatctc gataaaggcg cggcgcgcga atgcagctga
      781 tcacgtacgt ccctcgtgtt ccgttcaagg acggtgttat cgacctcaga ttaatgttta
      841 tcggccgact gttttcgtat ccgctcacca aacgcgtttt tgcattaaca ttgtatgtcg
      901 gcggatgttc tatatctaat ttgaataaat aaacgataac cgcgttggtt ttagagggca
      961 taataaaaga aatattgtta tcgtgttcgc cattagggca gtataaattg acgttcatgt
     1021 tggatattgt ttcagttgca agttgacact ggcggcgaca agatcgtgaa caaccaagtg
     1081 accatggatt acaaggatga cgatgacaag taactcgagc actaagtgat taacctcagg
     1141 ttatacatat attttgaatt taattaatta tacatatatt ttatattatt tttgtctttt
     1201 attatcgagg ggccgttgtt ggtgtggggt tttgcataga aataacaatg ggagttggcg
     1261 acgttgctgc gccaacacca cctcccttcc ctcctttcat catgtatctg tagataaaat
     1321 aaaatattaa acctaaaaac aagaccgcgc ctatcaacaa aatgataggc attaacttgc
     1381 cgctgacgct gtcactaacg ttggacgatt tgccgactaa accttcatcg cccagtaacc
     1441 aatctagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1501 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1561 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1621 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1681 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1741 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1801 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1861 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1921 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1981 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2041 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2101 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2161 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2221 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2281 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2341 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2401 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2461 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2521 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2581 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2701 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2761 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2821 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2881 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2941 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3001 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3061 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3241 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3301 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3361 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3421 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3481 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     3541 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     3601 tgaccatgat tacgaattcc cggg
//
