LOCUS       sequence-327078-        7246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Standard Gateway (attR1-attR2) vector for expression of His6-GST 
            fusions using the Bac-to-Bac insect cell system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7246)
  TITLE     Esposito Lab Plasmids
REFERENCE   2  (bases 1 to 7246)
  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: pDest-633
FEATURES             Location/Qualifiers
     source          1..7246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             146..163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             170..817
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="PILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFE
                     LGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGV
                     SRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYM
                     DPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     775..797
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             824..841
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     protein_bind    848..972
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(997..1002,1003..1020,1021..1027)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1081..1740
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1147..1166)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2061..2366
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2245..2264
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2407..2531)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2689..2823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2726..2745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2780..2799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2852..3017
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3201..3655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3288..3307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3497..3518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3681..3785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3786..3854,3855..4646)
                     /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(4004..4023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4817..5405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5306..5325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5559..5576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5710..5934
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6001..6534)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6723..6728,6729..6745,6746..6751))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(7114..7223)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcaccatgg gatcccatca ccatcaccat cacggcgccc ctatactagg
      181 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga
      241 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt
      301 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac
      361 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc
      421 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt
      481 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct
      541 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga
      601 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga
      661 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat
      721 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg
      781 gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctggttc cgcgtggatc
      841 ctcgatcaca agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa
      901 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
      961 ccagtcacta tggcggccgc attaggcacc ccaggcttta cactttatgc ttccggctcg
     1021 tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa ggaagctaaa
     1081 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1141 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1201 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     1261 cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt
     1321 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     1381 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     1441 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     1501 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     1561 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     1621 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     1681 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     1741 acgcgtggat ccggcttact aaaagccaga taacagtatg cgtatttgcg cgcaccggtg
     1801 ctagcgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga
     1861 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     1921 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     1981 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     2041 cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     2101 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     2161 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     2221 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     2281 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     2341 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     2401 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     2461 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
     2521 tacaaagtgg tgataattaa ttaagatcga gctagctaga agcttgtcga gaagtactag
     2581 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2641 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2701 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2761 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     2821 atctgatcac tgcttgagcc taggagatcc gaaccagata agtgaaatct agttccaaac
     2881 tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt
     2941 ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac
     3001 tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat caaacatcct
     3061 gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac agaatgaaaa
     3121 tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa cgcttatttg
     3181 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3241 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3301 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3361 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3421 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3481 gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3541 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3601 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3961 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4021 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5521 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5581 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5641 tagaccagcc gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg gatgccctgc
     5701 gtaagcgggt gtgggcggac aataaagtct taaactgaac aaaatagatc taaactatga
     5761 caataaagtc ttaaactaga cagaatagtt gtaaactgaa atcagtccag ttatgctgtg
     5821 aaaaagcata ctggactttt gttatggcta aagcaaactc ttcattttct gaagtgcaaa
     5881 ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg taaagactat attcgcggcg
     5941 ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg
     6001 ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt
     6061 tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc
     6121 accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct
     6181 ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa
     6241 cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag
     6301 cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg
     6361 ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga
     6421 tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa
     6481 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc
     6541 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     6601 gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
     6661 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
     6721 acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg
     6781 gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg
     6841 cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct
     6901 tctacggcaa ggtgctgtgc acggatctgc ccttgcttca ggagatcggt agacctcggc
     6961 cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc
     7021 tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac
     7081 tcgagatccc gggtgatcaa gtcttcgtcg agtgattgta aataaaatgt aatttacagt
     7141 atagtatttt aattaatata caaatgattt gataataatt cttatttaac tataatatat
     7201 tgtgttgggt tgaattaaag gtccgtatac tccggaatat taatag
//
