LOCUS       sequence-487565-        3746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect OpIE2 promoter expressing barnase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3746)
  AUTHORS   Nevard K, Ang JXD, Anderson MAE, Gonzalez E, Shackleford L, Alphey L
  TITLE     Harnessing the highly adaptable barnase-barstar system for genetic 
            biocontrol of Aedes aegypti.
  JOURNAL   Commun Biol. 2025 Aug 4;8(1):1154. doi: 10.1038/s42003-025-08588-6.
  PUBMED    40760041
REFERENCE   2  (bases 1 to 3746)
  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: "Commun 
            Biol."; date: "2025-08-4"; volume: "8(1):1154. doi"; pages: " 
            10.1038/s42003-025-08588-6"
COMMENT     Sequence Label: AGG1829 OpIE2-Bn
FEATURES             Location/Qualifiers
     source          1..3746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..548
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect 
                     cell expression"
     intron          581..713
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     696..715
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      780..789
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1245..1286
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1403..1524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1440..1459)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1494..1513
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1582..1598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1582..1598)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1595..1617)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1770..2358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1850..1869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2475..2499)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2599..3390,3391..3459))
                     /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     3222..3241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3460..3564)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3663..3685
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3677..3694
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3678..3694
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catgatgata aacaatgtat ggtgctaatg ttgcttcaac aacaattctg ttgaactgtg
       61 ttttcatgtt tgccaacaag cacctttata ctcggtggcc tccccaccac caactttttt
      121 gcactgcaaa aaaacacgct tttgcacgcg ggcccataca tagtacaaac tctacgtttc
      181 gtagactatt ttacataaat agtctacacc gttgtatacg ctccaaatac actaccacac
      241 attgaacctt tttgcagtgc aaaaaagtac gtgtcggcag tcacgtaggc cggccttatc
      301 gggtcgcgtc ctgtcacgta cgaatcacat tatcggaccg gacgagtgtt gtcttatcgt
      361 gacaggacgc cagcttcctg tgttgctaac cgcagccgga cgcaactcct tatcggaaca
      421 ggacgcgcct ccatatcagc cgcgcgttat ctcatgcgcg tgaccggaca cgaggcgccc
      481 gtcccgctta tcgcgcctat aaatacagcc cgcaacgatc tggtaaacac agttgaacag
      541 catctgttcc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag
      601 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
      661 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca
      721 ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag
      781 ccaccatggc ccaggttatc aacaccttcg atggagtggc cgattacctg cagacctacc
      841 acaagctgcc ggataactac atcaccaagt cggaagccca ggccctggga tgggtggcct
      901 cgaagggaaa cctggccgat gtggccccgg gaaagtcgat cggaggagat atcttctcga
      961 accgtgaagg aaagctgccg ggaaagtcgg gacgtacctg gcgtgaagta agaggagatc
     1021 ttgactaagt agatttatgg cgcaacgcta tcacaggaac ataactgaca taatctagac
     1081 gcgttactaa ctggcacctc ttcttttttt ttgatatcct gcaggccgat atcaactaca
     1141 cctcgggatt ccgtaactcg gatcgtatcc tgtactcgtc ggattggctg atctacaaga
     1201 ccaccgatca ctaccagacc ttcaccaaga tccgtggagc cggaggaaag ccgatcccga
     1261 acccgctgct gggactggat tcgacctaaa ctagttgatc ataatcagcc ataccacatt
     1321 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1381 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1441 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1501 gtccaaactc atcaatgtat cttacaggct aggtggaggc tcagtgatga taagtctgcg
     1561 atggtggatg catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcag
     1621 agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg gagttcagtt
     1681 cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1741 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1801 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1861 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1921 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1981 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2041 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2101 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2161 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2221 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2281 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2341 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctctattc
     2401 aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag ggggcttcaa atcgtcctcg
     2461 tgataccaat tcggagcctg cttttttgta caaacttgtt gataatggca attcaaggat
     2521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2701 gggcttacca tctggcccca gtgctgcaat gataccgcga gagccacgct caccggctcc
     2761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3361 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3421 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccagata cctgaaacaa
     3601 aacccatcgt acggccaagg aagtctccaa taactgtgat ccaccacaag cgccagggtt
     3661 ttcccagtca cgacgttgta aaacgacggc cagtcatgca taatccgcac gcatctggaa
     3721 taaggaagtg ccattccgcc tgacct
//
