LOCUS       sequence-467908-        5369 bp    DNA     circular SYN 15-APR-2025
DEFINITION  XanthoMoClo Golden Gate Backbone Plasmid, contains 
            Backbone_RK2-KanR_PartTypeBB and an sfGFP dropout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5369)
  AUTHORS   Soltysiak MPM, Ory ALH, Lee AD, Christophersen CE, Jalihal AP, 
            Springer M
  TITLE     XanthoMoClo─A Robust Modular Cloning Genetic Toolkit for the Genera 
            Xanthobacter and Roseixanthobacter
  JOURNAL   ACS Synthetic Biology. 2025. doi: 10.1021/acssynbio.4c00806.
REFERENCE   2  (bases 1 to 5369)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS 
            Synthetic Biology. 2025. doi: 10.1021/acssynbio.4c00806."
COMMENT     Sequence Label: pXTK-BB1 [Backbone_RK2-KanR_PartTypeBB]
FEATURES             Location/Qualifiers
     source          1..5369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     CDS             1074..1868
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1128..1147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1738..1757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    2054..2110
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     RBS             2293..2304
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2311..3024
                     /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      3036..3107
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3123..3150
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3199..3226)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3294..4442)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(4714..5085)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5118..5227
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agcccaatag
      721 cttcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg aacacgtaga
      781 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
      841 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
      901 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
      961 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     1021 gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     1081 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     1141 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     1201 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctccaagacg
     1261 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     1321 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     1381 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     1441 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     1501 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     1561 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcggatgccc gacggcgagg
     1621 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     1681 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     1741 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     1801 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     1861 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     1921 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     1981 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     2041 cccctgcggc cgcaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga
     2101 gctcgaattc gcgcggccgc ggcctaggcg gccaacgtga gaccgaaagt gaaacgtgat
     2161 ttcatgcgtc attttgaaca ttttgtaaat cttatttaat aatgtgtgcg gcaattcaca
     2221 tttaatttat gaatgttttc ttaacatcgc ggcaactcaa gaaacggcag gttcggatct
     2281 tagctactag agaaagagga gaaatactag atgcgtaaag gcgaagagct gttcactggt
     2341 gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt
     2401 ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact
     2461 ggtaaactgc cggttccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc
     2521 tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa
     2581 ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg
     2641 gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt
     2701 aaagaggacg gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt
     2761 tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac
     2821 gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat
     2881 ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat
     2941 ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg
     3001 catggtatgg atgaactgta caaatgacca ggcatcaaat aaaacgaaag gctcagtcga
     3061 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     3121 ctggctcacc ttcgggtggg cctttctgcg tttataggtc tctgagtcat atgcccacta
     3181 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgg
     3241 ttatcagaag agtgtaagcg ggttgactac cgaggaactg accccacaag gccctagcgt
     3301 ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct gcctcgcaac
     3361 tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga atccgatccg
     3421 cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga gctgaaatag
     3481 tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat gaatttcgtg
     3541 tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg gcaacgggac
     3601 gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga ttccaggtgc
     3661 ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg
     3721 gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag ctcgtagaac
     3781 gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac ctgctgccac
     3841 accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt cacgtccttg
     3901 ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg
     3961 gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc
     4021 gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac
     4081 gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt ttttcgcttc
     4141 ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc
     4201 tgttcgagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc agggggagcc
     4261 agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga gccgacggac
     4321 tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg
     4381 gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc
     4441 attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc ttattcctga
     4501 tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat gaagtcggtc
     4561 ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat
     4621 accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg
     4681 cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg ggcggaccac
     4741 gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca gcagggcgag
     4801 gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag
     4861 gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt gctcatagtc
     4921 cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggcct acaggctcat
     4981 gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc agtacacctt
     5041 gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct tgccctcatc
     5101 tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga gcaccgccag
     5161 gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat
     5221 cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa
     5281 tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa
     5341 gcagggttat gcagcggaaa agatccgtc
//
