LOCUS       sequence-456207-        4905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Moclo Baculo toolkit level 1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4905)
  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 4905)
  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.001
FEATURES             Location/Qualifiers
     source          1..4905
                     /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"
     RBS             2409..2420
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2427..3140
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3152..3223
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3239..3266
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      3288..3743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3375..3394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3585..3606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3770..3874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3875..3943,3944..4735)
                     /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(4093..4112)
                     /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 ttgagaccat gaaagtgaaa cgtgatttca
     2281 tgcgtcattt tgaacatttt gtaaatctta tttaataatg tgtgcggcaa ttcacattta
     2341 atttatgaat gttttcttaa catcgcggca actcaagaaa cggcaggttc ggatcttagc
     2401 tactagagaa agaggagaaa tactagatgc gtaaaggcga agagctgttc actggtgtcg
     2461 tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg
     2521 agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta
     2581 aactgccggt tccttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg
     2641 ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct
     2701 atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag
     2761 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
     2821 aggacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
     2881 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
     2941 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
     3001 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
     3061 acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg
     3121 gtatggatga actgtacaaa tgaccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3181 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     3241 ctcaccttcg ggtgggcctt tctgcgttta taagtcggtc tcagagtacg cgccctgtag
     3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3421 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3481 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3601 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3721 caaaatatta acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3781 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3841 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3901 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3961 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4021 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4081 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4141 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4201 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4261 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4321 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4381 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4441 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4501 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4561 tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt atcattgcag cactggggcc
     4621 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4681 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4741 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4801 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4861 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
