LOCUS       sequence-436698-        6889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector to generate recombinant baculovirus by site-specific 
            transposition in an E. coli host..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6889)
  TITLE     SGC-2024-Q1
REFERENCE   2  (bases 1 to 6889)
  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"
COMMENT     Sequence Label: pFHMSPN-avi-TEV-LIC V
FEATURES             Location/Qualifiers
     source          1..6889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..3533))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3885..3976
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3901..3918
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     sig_peptide     4032..4094
                     /codon_start=1
                     /product="signal sequence from honeybee melittin"
                     /label=melittin signal sequence
                     /translation="MKFLVNVALVFMVVYISYIYA"
     CDS             4110..4127
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4140..4184
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4206..4226
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4254..4287)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4302..4747
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(4748..4834,4835..6169)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     polyA_signal    6379..6513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6416..6435)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6470..6489
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6542..6707
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagca gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3961 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatctcg
     4021 gtccgaaaac catgaaattc ttagtcaacg ttgcccttgt ttttatggtc gtatacattt
     4081 cttacatcta tgcggccgct ccggaaatgc atcatcacca tcaccatgaa ttcatgagcg
     4141 gcctgaacga tatttttgaa gcgcagaaaa ttgaatggca tgaaggcagc gctggaggtt
     4201 caggtgaaaa cttgtatttc cagggcatta tgagttctcc tcctgaaaga tccataactt
     4261 cgtatagcat acattatacg aagttatgcg gccgcgacgt ccacatatac ctgccgttca
     4321 ctattattta gtgaaatgag atattatgat attttctgaa ttgtgattaa aaaggcaact
     4381 ttatgcccat gcaacagaaa ctataaaaaa tacagagaat gaaaagaaac agatagattt
     4441 tttagttctt taggcccgta gtctgcaaat ccttttatga ttttctatca aacaaaagag
     4501 gaaaatagac cagttgcaat ccaaacgaga gtctaataga atgaggtcga aaagtaaatc
     4561 gcgcgggttt gttactgata aagcaggcaa gacctaaaat gtgtaaaggg caaagtgtat
     4621 actttggcgt caccccttac atattttagg tcttttttta ttgtgcgtaa ctaacttgcc
     4681 atcttcaaac aggagggctg gaagaagcag accgctaaca cagtacataa aaaaggagac
     4741 atgaacgatg aacatcaaaa agtttgcaaa acaagcaaca gtattaacct ttactaccgc
     4801 actgctggca ggaggcgcaa ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa
     4861 ggaaacatac ggcatttccc atattacacg ccatgatatg ctgcaaatcc ctgaacagca
     4921 aaaaaatgaa aaatataaag ttcctgagtt cgattcgtcc acaattaaaa atatctcttc
     4981 tgcaaaaggc ctggacgttt gggacagctg gccattacaa aacactgacg gcactgtcgc
     5041 aaactatcac ggctaccaca tcgtctttgc attagccgga gatcctaaaa atgcggatga
     5101 cacatcgatt tacatgttct atcaaaaagt cggcgaaact tctattgaca gctggaaaaa
     5161 cgctggccgc gtctttaaag acagcgacaa attcgatgca aatgattcta tcctaaaaga
     5221 ccaaacacaa gaatggtcag gttcagccac atttacatct gacggaaaaa tccgtttatt
     5281 ctacactgat ttctccggta aacattacgg caaacaaaca ctgacaactg cacaagttaa
     5341 cgtatcagca tcagacagct ctttgaacat caacggtgta gaggattata aatcaatctt
     5401 tgacggtgac ggaaaaacgt atcaaaatgt acagcagttc atcgatgaag gcaactacag
     5461 ctcaggcgac aaccatacgc tgagagatcc tcactacgta gaagataaag gccacaaata
     5521 cttagtattt gaagcaaaca ctggaactga agatggctac caaggcgaag aatctttatt
     5581 taacaaagca tactatggca aaagcacatc attcttccgt caagaaagtc aaaaacttct
     5641 gcaaagcgat aaaaaacgca cggctgagtt agcaaacggc gctctcggta tgattgagct
     5701 aaacgatgat tacacactga aaaaagtgat gaaaccgctg attgcatcta acacagtaac
     5761 agatgaaatt gaacgcgcga acgtctttaa aatgaacggc aaatggtacc tgttcactga
     5821 ctcccgcgga tcaaaaatga cgattgacgg cattacgtct aacgatattt acatgcttgg
     5881 ttatgtttct aattctttaa ctggcccata caagccgctg aacaaaactg gccttgtgtt
     5941 aaaaatggat cttgatccta acgatgtaac ctttacttac tcacacttcg ctgtacctca
     6001 agcgaaagga aacaatgtcg tgattacaag ctatatgaca aacagaggat tctacgcaga
     6061 caaacaatca acgtttgcgc ctagcttcct gctgaacatc aaaggcaaga aaacatctgt
     6121 tgtcaaagac agcatccttg aacaaggaca attaacagtt aacaaataaa aacgcaaaag
     6181 aaaatgccga tatcctattg gcattgacgt caggtggcac ttttcgagga gatcatgcac
     6241 atgatgacga agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta
     6301 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     6361 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6421 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6481 aaactcatca atgtatctta tcatgtctgg atctgatcac tgatatcgcc taggagatcc
     6541 gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt tcgtattagc
     6601 ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata atccttaaaa
     6661 actccatttc cacccctccc agttcccaac tattttgtcc gcccacagcg gggcattttt
     6721 cttcctgtta tgtttttaat caaacatcct gccaactcca tgtgacaaac cgtcatcttc
     6781 ggctactttt tctctgtcac agaatgaaaa tttttctgtc atctcttcgt tattaatgtt
     6841 tgtaattgac tgaatatcaa cgcttatttg cagcctgaat ggcgaatgg
//
