LOCUS       sequence-456232-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Moclo Baculo toolkit level 2 backbone (biGBac compatible).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5860)
  AUTHORS   Lai Z, Flanigan SF, Boudes M, Davidovich C
  TITLE     Modular cloning of multigene vectors for the baculovirus system and 
            yeast.
  JOURNAL   J Mol Biol. 2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943.
  PUBMED    39814169
REFERENCE   2  (bases 1 to 5860)
  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: "J Mol Biol.
            2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943."
COMMENT     Sequence Label: pMB.BIG1d
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  892..1116
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1183..1716)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1905..1910,1911..1927,1928..1933))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     mobile_element  2716..2881
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             3405..4196
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4243..4698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4330..4349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4540..4561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4725..4829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4830..4898,4899..5690)
                     /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(5048..5067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcagaccag ccgcgtaacc
      841 tggcaaaatc ggttacggtt gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg
      901 acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag tcttaaacta
      961 gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt
     1021 ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag
     1081 aggggcgtgg ccaagggcat ggtaaagact atattcgcgg cgttgtgaca atttaccgaa
     1141 caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg
     1201 gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc
     1261 gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc
     1321 atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac
     1381 tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc
     1441 cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact
     1501 acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc
     1561 gccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc
     1621 gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact
     1681 gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac
     1741 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa
     1801 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     1861 aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac
     1921 aggcttatgt caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa
     1981 ccttgggcag cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt
     2041 cggactccac gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt
     2101 gcacggatct gccctggctt caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg
     2161 tggtgctgac cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt
     2221 tgttcgccca ggactctagc tatagttccc tgaattcgca tctagagttt aaacctaatg
     2281 atgcctgatg tttctgatga gacgtggtag agccacaaac agccggtaca agcaacgatc
     2341 tccaggacca tctgaatcat gcgcggatga cacgaactca cgacggcgat cacagacatt
     2401 aacccacagt acagacactg cgacaacgtg gcaattcgtc gcaatacaac gactagagat
     2461 ctatgtgagg atcctaactc gagatcgagc tggaaatctg ctcgtcagtg gtgctcacac
     2521 tgacgaatca tgtacagatc ataccgatga ctgcctggcg actcacaact aagcaagaca
     2581 gccggaacca gcgccggcga acaccactgc atatatggca tatcacaaca gtccacgtct
     2641 caagcaaaac ggttcacata gcttagttta aacgttacag agatgttacg aaccactagt
     2701 gcactgcagt acatcaacca gataagtgaa atctagttcc aaactatttt gtcattttta
     2761 attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta
     2821 aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac
     2881 agcggggcat ttttcttcct gttatgtttt taatcaaaca tcctgccaac tccatgtgac
     2941 aaaccgtcat cttcggctac tttttctctg tcacagaatg aaaatttttc tgtcatctct
     3001 tcgttattaa tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaactgggag
     3061 gcagaataaa tgatcatatc gtcaattatt acctccacgg ggagagcctg agcaaactgg
     3121 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaagt
     3181 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     3241 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     3301 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     3361 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg gaagcggtga
     3421 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     3481 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     3541 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     3601 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     3661 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     3721 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     3781 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     3841 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     3901 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     3961 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     4021 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     4081 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     4141 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc
     4201 taacaattcg ttcaagccga cggattggcg aatgggggga gtacgcgccc tgtagcggcg
     4261 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4321 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4381 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4441 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4501 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4561 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4621 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4681 tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4741 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4801 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4861 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4921 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4981 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5041 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5101 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5161 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5221 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5281 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5341 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5401 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5461 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5521 ataaatctgg agccggtgag cgtggttctc gcggtatcat tgcagcactg gggccagatg
     5581 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5641 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5701 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5761 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5821 actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
