LOCUS       sequence-469235-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Moclo Baculo toolkit level 1 backbone with ConL4/ConR5 and Tn7L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  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 5623)
  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.Pos5
FEATURES             Location/Qualifiers
     source          1..5623
                     /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             2601..2612
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2619..3332
                     /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      3344..3415
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3431..3458
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     mobile_element  3651..3816
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4006..4461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4093..4112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4303..4324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4488..4592
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4593..4661,4662..5453)
                     /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(4811..4830)
                     /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 tctagagcct gcactagacg
     2281 aactaggcaa gatgcgtcca atccgtctaa acatggtgac aacgctggac agatgacgta
     2341 acaccgagcc acatcctgaa atcgaggcag gctaaccgaa accgtgacaa tgcaaagaga
     2401 cagcctgacc gtctcaggta gactacccat gagtcacaat gaacgtgaga ccgaaagtga
     2461 aacgtgattt catgcgtcat tttgaacatt ttgtaaatct tatttaataa tgtgtgcggc
     2521 aattcacatt taatttatga atgttttctt aacatcgcgg caactcaaga aacggcaggt
     2581 tcggatctta gctactagag aaagaggaga aatactagat gcgtaaaggc gaagagctgt
     2641 tcactggtgt cgtccctatt ctggtggaac tggatggtga tgtcaacggt cataagtttt
     2701 ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg aagttcatct
     2761 gtactactgg taaactgccg gttccttggc cgactctggt aacgacgctg acttatggtg
     2821 ttcagtgctt tgctcgttat ccggaccata tgaagcagca tgacttcttc aagtccgcca
     2881 tgccggaagg ctatgtgcag gaacgcacga tttcctttaa ggatgacggc acgtacaaaa
     2941 cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca
     3001 ttgactttaa agaggacggc aatatcctgg gccataagct ggaatacaat tttaacagcc
     3061 acaatgttta catcaccgcc gataaacaaa aaaatggcat taaagcgaat tttaaaattc
     3121 gccacaacgt ggaggatggc agcgtgcagc tggctgatca ctaccagcaa aacactccaa
     3181 tcggtgatgg tcctgttctg ctgccagaca atcactatct gagcacgcaa agcgttctgt
     3241 ctaaagatcc gaacgagaaa cgcgatcata tggttctgct ggagttcgta accgcagcgg
     3301 gcatcacgca tggtatggat gaactgtaca aatgaccagg catcaaataa aacgaaaggc
     3361 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     3421 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tataggtctc agctgaagtt
     3481 gagacgacgc acaaggtcag ggcactcatg cgacaatcaa ctcgatgcat gatccgcacc
     3541 attgtcgagg ggccagcgtc aatagtgccg atgaccacag acccggttaa gacatagccg
     3601 aatggagccg cgccgaccac agaatgataa ctagtgcact gcagtacatc aaccagataa
     3661 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
     3721 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
     3781 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
     3841 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
     3901 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
     3961 gaatatcaac gcttatttgc agcctgaatg gcgaatgggg ggagtacgcg ccctgtagcg
     4021 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4081 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4141 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4201 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4261 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4321 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4381 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4441 aaatattaac gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4501 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4561 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4621 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4681 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4741 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4801 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4861 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4921 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4981 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5041 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5101 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5161 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5221 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5281 ctgataaatc tggagccggt gagcgtggtt ctcgcggtat cattgcagca ctggggccag
     5341 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5401 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5461 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5521 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5581 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
