LOCUS       sequence-467922-        8544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  XanthoMoClo Golden Gate Backbone Plasmid, contains 
            Backbone_repABC-pMlb-KanR_PartTypeBB and an sfGFP dropout .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8544)
  AUTHORS   Soltysiak MPM, Ory ALH, Lee AD, Christophersen CE, Jalihal AP, 
            Springer M
  TITLE     XanthoMoClo horizontal line A Robust Modular Cloning Genetic Toolkit
            for the Genera Xanthobacter and Roseixanthobacter.
  JOURNAL   ACS Synth Biol. 2025 Apr 18;14(4):1173-1190. doi: 
            10.1021/acssynbio.4c00806. Epub 2025 Mar 13.
  PUBMED    40080684
REFERENCE   2  (bases 1 to 8544)
  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"; doi: 
            "10.1021/acssynbio.4c00806"; journalName: "ACS Synth Biol"; date: 
            "2025-04-18- 18"; volume: "14"; issue: "4"; pages: "1173-1190"
COMMENT     Sequence Label: pXTK-BB3 [Backbone_repABC-pMlb-KanR_PartTypeBB]
FEATURES             Location/Qualifiers
     source          1..8544
                     /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"
     CDS             964..1758
                     /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(1018..1037)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1628..1647
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    6640..6696
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     RBS             6879..6890
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6897..7610
                     /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      7622..7693
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7709..7736
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     oriT            7889..7998
                     /note="incP origin of transfer"
     CDS             8031..8402
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct 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 cgcgcaatag cttcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg
      661 aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
      721 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
      781 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
      841 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
      901 ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt
      961 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     1021 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     1081 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     1141 ctccaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     1201 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     1261 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     1321 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     1381 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     1441 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcggatgccc
     1501 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     1561 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     1621 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     1681 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     1741 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     1801 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     1861 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     1921 tcgcccaccc cccctggcgt tcgtcactcg tttggccatg cggaatctcc agccggaaca
     1981 ccccctcgtc gcccggctgg cggtccgcgc gggattggct tgcgatttca ctttgtcagc
     2041 aagcggtgct gtttgcgcat cgtgacgggt tggtaccgct ggtctattgc cggcccctac
     2101 cccgatagat ttcatccgca ttgacacgac gatgtgttga agccgggaaa tgtgacgagc
     2161 cgtcttcagg agtctttggc ctcggtctgg cgatgctggc cgtcagcatg cgtctcaaca
     2221 tagcggtctt ggcaacacca ccgcgtacct acgatctgaa actttgctcc gccgaatggt
     2281 cgcgtctggt gcgcgcaaat ccatgttttc aatatcgttc tgcgattttt ttactgcggt
     2341 aaatttctgg atgcggtcag tgcgattggc cgcgctctgg ccagcgttaa ccttttgtta
     2401 acccaaaact tcttgccggt aagtccgaat cggagcttaa tggcgcttgt tgtgggcatt
     2461 gcttacaaaa agcgccaatt caggtttcga cgagaatgaa tgtctccatc cggctgccac
     2521 gtttcgagtt cgatgacaaa atcctcgaac agggcgccga gatttcccgg cggttgaacc
     2581 agcttcggat ggagaaattt ccgcccaatg cgacgaagac gctacgcaca ttctcgatgg
     2641 cggaggtcgc tcactacctc ggggtttcgc agaacaattt gaagcgtctc catctcgaag
     2701 gcaaggggcc gttgcctgac caaagtcctt ccggccgtcg ctcctacacg gctacacaga
     2761 tgctcgagct caggcagttc ctggaccagc atggcaggac tgaagtcaag aagtacgttc
     2821 cgcaccgcaa gcttggcgat aagctccagg tggttgcggt ggtcaatttc aaaggcggca
     2881 gcggcaagac gaccacggca gcccacctcg cccagcacct ggcgctgact ggtcatcggg
     2941 ttcttgcgat cgatcttgat ccgcaagctt cgctgtctgc tctgcacggc ttccagcccg
     3001 agctcgatcg caacctctcg ctctatgaag cgatcaggta tgacgacgaa cgcagatcga
     3061 ttgcggatgt gatcataccg acaaatttcc ccggcctcga catcattccc gccaatctcg
     3121 agctgcagga atacgagtat gacacgccgc tcgccatgca agacaaaacc tccagtgccg
     3181 gccgccagtt cttcacgcgc atggccaaag cgttgtcgga ggttgacgat cggtacgatg
     3241 tcgtcgttgt cgattgcccg ccccagcttg gctatctgac tttgacggca atgtcggcgg
     3301 cgacatcggt tctcattacg gtccatccgc agatgctcga cctgatgtcg atgagccagt
     3361 ttcttctgat gcttggcggc atattgaaga ccatcaaggc cgccggcgcc gcgatcgaac
     3421 ttgactggtt tcgctacctg atcacgcgct acgaaccgac cgatattccg caggcgcaga
     3481 tggtgggctt catgcagtcc atgcttgccg gccagatcct ggaaaacccg atgttgaagt
     3541 cgacggcgat ttccgatgca ggcctcacca agcaaacact ttacgaggtg gagaaatcgg
     3601 ccttcacccg ctcgacatat gaccgcgctt tggaatcgct cgacgcggtg aacgccgaga
     3661 tcgtcacact catccatcgt gcgtggggtc ggtcatgacg gcgaacttta cggcagtaaa
     3721 aaccggctat tttgctcgtg gaatcagatc gctagccgct cacggaggcg gttgaatggc
     3781 ccgtaaggat attttcggca gtgtgatggg acccgtcgct gcggtggccg ataacaagga
     3841 tgtatcgggg tatgccgttc gtggtgcctc ccgctcgatc atgcaatcct tcgaggagct
     3901 ctccaagagt tccgtcgtcg atctcgatcc tgccctggtt gacacctcgt tcgtctcgga
     3961 ccgcctggac gaagacgatc aggcgtatca ggagttgctg tcggcaattc gcgagagagg
     4021 tcaagattct ccgattctgg tcagaccgca tccggaggtt tcaggacgct tccagacagt
     4081 attcggccat cgccgcgtca aggttgcccg ccagctggga aggccggtcc gcgccgtcgt
     4141 caagaagatg ggcgacgagg accatgtcat cgcccagggc caggagaatg ccgccagggc
     4201 caatctgtcc ttcatcgaga ggaccctgtt tgccgatcgc attctcaagc gcggccacac
     4261 ctcggacacg gtgaagtccg ctttggcgct cgatgacacg accttttcga agatgcgctc
     4321 cgtgaccggc aatatccccg agccagtcat tgtggctgtt ggctcggcca aggccgtcgg
     4381 acgggacagg tggtggcaac tggccaagct catggagcat cctgggaacg cgggcagggc
     4441 ggccgactat gtgaagagcg ctgatttcgc ggagccaagc agtgacgcac gatttgcgct
     4501 tcttttcgac tacctgaacg ctccggcgaa acgttcgccg gcagcgccga agacacgaga
     4561 aaaggcctgg gcgccgcaag acaggtcggt gcgggccaag atcaccgaca ccggaaaagt
     4621 gttcacgctt gccctgaaag caagggaagc ctcgccgttc ggtgcgttca tcaccgaacg
     4681 gctggacgag cttttcgaag cgttccggca atcggaaatg tctaagagaa caggagactg
     4741 aaaccgcaaa agaaaaaggc ccccgaacgt cgccgtcgcg gaagcccttc tcgtctgtag
     4801 caagctgaga atcgcacttc cccgaatcac tgtcaagagt cgtggcgcca tttcggcgag
     4861 caggtttctt ttgccttcgt gaaaggtgag ggaaatggag agtgggtttg cgacgacgcc
     4921 ttttgggcgg cggccgatga cgcttgcctt ggtgaaggcc aatgccgcgt cgcggcagat
     4981 tccgaaaggg gccgccgttg agaaatggca ggtctaccgg tggctatgcg agggcaagtc
     5041 gatcgtcggc gttggcgatc gtgccctggc ggtgctgagc gggctgttga cgttttatcc
     5101 tgacgaccag atcagcgagg aaaacggctt gatggtcttc ccgtcgaacg cacagctgtc
     5161 tttgcgggcg cacggcatgg ccgattcgac gctgagaagg aacctggccg agctggtgag
     5221 ttgcggcctg atcattcgcc gcgatagtcc gaacggcaaa agattcgctc gtaaggggca
     5281 gggtggtgct gtcatggagg tcttcgggtt ctcgttggcg cctctgctgg cacgggcggc
     5341 ggagtttcag cgcgccgccg agcaggtccg cgccgagtgc caggcgctga aattgatgcg
     5401 ggagcgcatc acgctgcatc gcagggacat tcagaagctg gtcgaggcgg cgatcgagga
     5461 aggggttccg ggcgactggg gcggcgtctg gaggcggttt cggtccatcg tggatgcaat
     5521 tccgcgccgg gcggggctgt cacagcttga agggatcgtt gccgagctgg tcgtgctcca
     5581 cgatgaagtg gataggatgc tggaaagctt cacaagttcc acgaatccta gcggcaatga
     5641 gtcccaaaat gagcggcagc aatctgattc aaacacagac tccatttttg tatttgaacc
     5701 ggctttcgaa aaaagcaggg cggccgttga gccaagaccg gcaccggacg agcggttgaa
     5761 aagctatccg attggactgg tgctgaaggc ctgccccgag tttgcgggct atgctgtcaa
     5821 cgggatcgcc agctggcgcg atttgatgct caccgcggcc caggtgcgcg gctaccttgg
     5881 ggtttcaccg tcggcctatg aggaggcttg ccatgtgatg ggtcaggaag tcgccgcgat
     5941 tgtgattgcg tgtattctgc aacgggcaca gcacatcaat tcggccggtg gctatctgcg
     6001 cgtgctgacg gaaaaggcga gggcagggga gttctcggtc ggaccgatgc tgatggccgc
     6061 gctcaaggcc aatggcccgg caacgaggat gacgggatga ttgaggtaca ggttcgtggg
     6121 gcaggttgct cggcgctcta ggtgacggca tgcggccacc taccggcggt gggctcccgg
     6181 gcttgaaagc tgttcaggaa tgccatatct tttgcacaag aaacatgact cctgatatgg
     6241 tgaacactgt gggccatgca ctcaaggcag ccgagcagaa tgggccgctc gtcttgtcct
     6301 atatggatcg gaacgggaag attgcgatcg agcaggtcgc ggacggtttt ggcatgtcca
     6361 agggtcagtt ggcccagacc gccggtcttg cgcgtgagac actctatcgc tcagagcgca
     6421 gcgctgcggc aaagacccag gggcgtctgc gcgaaatgct cgagatcatc agccgcgtga
     6481 gcgattgggc gggcggcaag gaacaggcga tggcctggta tcgggctcag ccgcttccgg
     6541 catttggcgg acgcactgcc gaagcgctgg tgaaggatgg caaggccgcc gcggtccgtg
     6601 attatctaga tcacatggca gtcggtgtgg cgcggccgca agcttgcatg cctgcaggtc
     6661 gactctagag gatccccggg taccgagctc gaattcgcgc ggccgcggcc taggcggcca
     6721 acgtgagacc gaaagtgaaa cgtgatttca tgcgtcattt tgaacatttt gtaaatctta
     6781 tttaataatg tgtgcggcaa ttcacattta atttatgaat gttttcttaa catcgcggca
     6841 actcaagaaa cggcaggttc ggatcttagc tactagagaa agaggagaaa tactagatgc
     6901 gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg
     6961 tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac
     7021 tgacgctgaa gttcatctgt actactggta aactgccggt tccttggccg actctggtaa
     7081 cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg
     7141 acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg
     7201 atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc
     7261 gcattgagct gaaaggcatt gactttaaag aggacggcaa tatcctgggc cataagctgg
     7321 aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta
     7381 aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact
     7441 accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga
     7501 gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg
     7561 agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa tgaccaggca
     7621 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     7681 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     7741 taggtctctg agtatcttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt
     7801 cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact
     7861 ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga
     7921 gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg
     7981 ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg atggctgatg
     8041 aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt gtactgcctt ccagacgaac
     8101 gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg
     8161 ccgtcggcca gggctacaaa atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg
     8221 cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg
     8281 acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc cctgctggcg aagatcgaag
     8341 agaagcagga cgagcttggc aaggtcatga tgggcgtggt ccgcccgagg gcagagccat
     8401 gactttttta gccgctaaaa cggccggggg gtgcgcgtga ttgccaagca cgtccccatg
     8461 cgctccatca agaagaggca cttcgagctg taagtacatc accgacgagc aaggcaagac
     8521 gatcccgaag atcaaaggat cttc
//
