LOCUS       sequence-322112-        7830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Standard Gateway (attR1-attR2) vector for expression of 
            maltose-binding protein (MBP) 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 7830)
  TITLE     Esposito Lab Plasmids
REFERENCE   2  (bases 1 to 7830)
  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-795
FEATURES             Location/Qualifiers
     source          1..7830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..119
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     44..61
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             176..1273
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1130..1139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1247..1270
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1292..1416
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1441..1446,1447..1464,1465..1471)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1525..2184
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1591..1610)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2526..2831
                     /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     2710..2729
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2872..2996)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3141..3275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3178..3197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3232..3251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3304..3469
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3653..4107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3740..3759)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3949..3970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4133..4237
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4238..4306,4307..5098)
                     /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(4456..4475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5269..5857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5758..5777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6011..6028
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6162..6386
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6453..6986)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7175..7180,7181..7197,7198..7203))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    7711..7759
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     7784..7803
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 tcgaggtata ctccggaata ttaatagatc atggagataa ttaaaatgat aaccatctcg
       61 caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa cctataaata
      121 ttccggatta ttcataccgt cccaccatcg ggcgcggatc accatgggat ccggcaaaat
      181 cgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga
      241 agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa
      301 actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg
      361 ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga
      421 caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa
      481 gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct
      541 gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa
      601 aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc
      661 tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg
      721 cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa
      781 acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac
      841 agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta
      901 tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct
      961 gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa
     1021 ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt
     1081 agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga
     1141 aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc
     1201 cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa
     1261 agacgcgcag actaattcgg gatcctcgat cacaagtttg tacaaaaaag ctgaacgaga
     1321 aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac
     1381 ataatactgt aaaacacaac atatccagtc actatggcgg ccgcattagg caccccaggc
     1441 tttacacttt atgcttccgg ctcgtataat gtgtggattt tgagttagga tccgtcgaga
     1501 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     1561 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     1621 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     1681 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     1741 ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     1801 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     1861 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     1921 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     1981 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     2041 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     2101 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     2161 gatgagtggc agggcggggc gtaaacgcgt ggatccggct tactaaaagc cagataacag
     2221 tatgcgtatt tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga
     2281 agtatgtcaa aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca
     2341 gctatcagtt gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat
     2401 gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat
     2461 ggctgaggtc gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg
     2521 gtgaaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg
     2581 atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg
     2641 cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg
     2701 aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag
     2761 aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct
     2821 ggggaatata aatgtcaggc tcccttatac acagccagtc tgcaggtcga ccatagtgac
     2881 tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat
     2941 atattgatat ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggtgatc
     3001 gagctagcta gaagcttgtc gagaagtact agaggatcat aatcagccat accacatttg
     3061 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3121 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3181 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3241 ccaaactcat caatgtatct tatcatgtct ggatctgatc actgcttgag cctaggagat
     3301 ccgaaccaga taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta
     3361 gcttacgacg ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa
     3421 aaactccatt tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt
     3481 ttcttcctgt tatgttttta atcaaacatc ctgccaactc catgtgacaa accgtcatct
     3541 tcggctactt tttctctgtc acagaatgaa aatttttctg tcatctcttc gttattaatg
     3601 tttgtaattg actgaatatc aacgcttatt tgcagcctga atggcgaatg ggacgcgccc
     3661 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3721 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3781 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3841 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3901 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tcttaatagt ggactcttgt
     3961 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4021 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4081 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     4141 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4201 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4261 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4321 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4381 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4441 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4501 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4561 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4621 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4681 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4741 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4801 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4861 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4921 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4981 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5041 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5101 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5161 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5221 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5281 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5341 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5401 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5461 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5521 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5581 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5641 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5701 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5761 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5821 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5881 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5941 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6001 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     6061 ccttacgcat ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc
     6121 ggttacggtt gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt
     6181 cttaaactga acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag
     6241 ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc
     6301 taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg
     6361 ccaagggcat ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg
     6421 gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca
     6481 aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca
     6541 ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg
     6601 agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca
     6661 tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg
     6721 ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag
     6781 ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca
     6841 cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat
     6901 gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc
     6961 gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     7021 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata
     7081 acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga
     7141 ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt
     7201 caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag
     7261 cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac
     7321 gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct
     7381 gcccttgctt caggagatcg gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac
     7441 cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca
     7501 ggactctagc tatagttcta gtggttggct acgtacccgt agtggctatg gcagggcttg
     7561 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg
     7621 aaaaggaaga aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt
     7681 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta
     7741 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     7801 tcagttagcc tcccccatct cccggtaccc
//
