LOCUS       sequence-13854-e        6472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6472)
  TITLE     5-series pFastBac Dual LIC vectors (for insect cell expression)
REFERENCE   2  (bases 1 to 6472)
  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: pFastBac Dual His6 MBP N10 TEV LIC cloning vec...
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     399..420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        583..687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(688..756,757..1548)
                     /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(906..925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1719..2307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2208..2227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2461..2478
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2610..2834
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2901..3434)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDVDPSTAT"
     promoter        complement(join(3623..3628,3629..3645,3646..3651))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4160..4208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4233..4252
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4348..4457)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4481..4572
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4497..4514
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4634..4651
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4661..5761
                     /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="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     5735..5758
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             5816..5836
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    6062..6196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6099..6118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6153..6172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6225..6390
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaact
      541 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      661 cttcaatact attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      721 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      781 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      841 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
      901 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
      961 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1021 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1081 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1141 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1201 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1261 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1321 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1381 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1441 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1501 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1561 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1621 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1681 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1741 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1801 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1861 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1921 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1981 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2041 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2101 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2161 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2221 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2281 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2341 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2401 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2461 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     2521 ctgtgcggta tttcacaccg cagaccagcc gcgtaacctg gcaaaatcgg ttacggttga
     2581 gtaataaatg gatgccctgc gtaagcgggt gtgggcggac aataaagtct taaactgaac
     2641 aaaatagatc taaactatga caataaagtc ttaaactaga cagaatagtt gtaaactgaa
     2701 atcagtccag ttatgctgtg aaaaagcata ctggactttt gttatggcta aagcaaactc
     2761 ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg
     2821 taaagactat attcgcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg
     2881 atctcggctt gaacgaattg ttaggtggcg gtacttgggt cgacatcaaa gtgcatcact
     2941 tcttcccgta tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct
     3001 tgcacgtaga tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc
     3061 gcggtggcaa tgccctgcct ccggtgctcg ccggagactg cgagatcata gatatagatc
     3121 tcactacgcg gctgctcaaa cctgggcaga acgtaagccg cgagagcgcc aacaaccgct
     3181 tcttggtcga aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa
     3241 tcggagtccg gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga
     3301 tcaagagcag cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg
     3361 cccatacttg agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg
     3421 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     3481 ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa cagtcataac aagccatgaa
     3541 aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga
     3601 gcgcatacgc tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggttcg
     3661 tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga
     3721 ggcatttctg tcctggctgg cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc
     3781 attggcggcc ttgctgttct tctacggcaa ggtgctgtgc acggatctgc cctggcttca
     3841 ggagatcggt agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt
     3901 ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg actctagcta
     3961 tagttctagt ggttggccta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4261 cccccatctc ccggtaccgc atgccttatg gagttgggat atctatatct ccttcttaaa
     4321 gcccgggtga tcaagtcttc gtcgagtgat tgtaaataaa atgtaattta cagtatagta
     4381 ttttaattaa tatacaaatg atttgataat aattcttatt taactataat atattgtgtt
     4441 gggttgaatt aaaggtccgt atactccgga ctattaatag atcatggaga taattaaaat
     4501 gataaccatc tcgcaaataa ataagtattt tactgttttc gtaacagttt tgtaataaaa
     4561 aaacctataa ctattccgga ttattcatac cgtcccacca tcgggcgcgg atcccggtcc
     4621 gatgggttct tctcaccatc accatcacca tggttcttct atgaaaatcg aagaaggtaa
     4681 actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa
     4741 attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa
     4801 attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg
     4861 ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca
     4921 ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta
     4981 cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc
     5041 aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc
     5101 gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg
     5161 ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc
     5221 tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc
     5281 agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat
     5341 caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt
     5401 actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat
     5461 taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac
     5521 tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc
     5581 ttacgaggaa gagttggcga aagatccacg tattgccgcc actatggaaa acgcccagaa
     5641 aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc
     5701 ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac
     5761 taattcgagc tcgaacaaca acaacaataa caataacaac aacctcggga tcgaggaaaa
     5821 cctgtacttc caatccaata ttggaagtgg ataacggatc cgaattcaaa ggcctacgtc
     5881 gacgagctca ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg acaagcttgt
     5941 cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     6001 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     6061 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6121 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6181 ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag ataagtgaaa
     6241 tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca
     6301 gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct
     6361 cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt
     6421 aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact tt
//
