LOCUS       sequence-467909-        7488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  XanthoMoClo Golden Gate Backbone Plasmid, contains 
            Backbone_RK2-KanR-TetR_PartTypeBB and an sfGFP dropout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7488)
  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 7488)
  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-BB2 [Backbone_RK2-KanR-TetR_PartTypeBB]
FEATURES             Location/Qualifiers
     source          1..7488
                     /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"
     CDS             complement(2138..2788)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             2894..4093
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     RBS             4412..4423
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4430..5143
                     /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      5155..5226
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5242..5269
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5318..5345)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(5413..6561)
                     /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(6833..7204)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7237..7346
                     /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 gcctggctca atcgtcaccc tttctcggtc
     2161 cttcaacgtt cctgacaacg agcctccttt tcgccaatcc atcgacaatc accgcgagtc
     2221 cctgctcgaa cgctgcgtcc ggaccggctt cgtcgaaggc gtctatcgcg gcccgcaaca
     2281 gcggcgagag cggagcctgt tcaacggtgc cgccgcgctc gccggcatcg ctgtcgccgg
     2341 cctgctcctc aagcacggcc ccaacagtga agtagctgat tgtcatcagc gcattgacgg
     2401 cgtccccggc cgaaaaaccc gcctcgcaga ggaagcgaag ctgcgcgtcg gccgtttcca
     2461 tctgcggtgc gcccggtcgc gtgccggcat ggatgcgcgc gccatcgcgg taggcgagca
     2521 gcgcctgcct gaagctgcgg gcattcccga tcagaaatga gcgccagtcg tcgtcggctc
     2581 tcggcaccga atgcgtatga ttctccgcca gcatggcttc ggccagtgcg tcgagcagcg
     2641 cccgcttgtt cctgaagtgc cagtaaagcg ccggctgctg aacccccaac cgttccgcca
     2701 gtttgcgtgt cgtcagaccg tctacgccga cctcgttcaa caggtccagg gcggcacgga
     2761 tcactgtatt cggctgcaac tttgtcatgc ttgacacttt atcactgata aacataatat
     2821 gtccaccaac ttatcagtga taaagaatcc gcgcgttcaa tcggaccagc ggaggctggt
     2881 ccggaggcca gacgtgaaac ccaacatacc cctgatcgta attctgagca ctgtcgcgct
     2941 cgacgctgtc ggcatcggcc tgattatgcc ggtgctgccg ggcctcctgc gcgatctggt
     3001 tcactcgaac gacgtcaccg cccactatgg cattctgctg gcgctgtatg cgttggtgca
     3061 atttgcctgc gcacctgtgc tgggcgcgct gtcggatcgt ttcgggcggc ggccaatctt
     3121 gctcgtctcg ctggccggcg ccactgtcga ctacgccatc atggcgacag cgcctttcct
     3181 ttgggttctc tatatcgggc ggatcgtggc cggcatcacc ggggcgactg gggcggtagc
     3241 cggcgcttat attgccgata tcactgatgg cgatgagcgc gcgcggcact tcggcttcat
     3301 gagcgcctgt ttcgggttcg ggatggtcgc gggacctgtg ctcggtgggc tgatgggcgg
     3361 tttctccccc cacgctccgt tcttcgccgc ggcagccttg aacggcctca atttcctgac
     3421 gggctgtttc cttttgccgg agtcgcacaa aggcgaacgc cggccgttac gccgggaggc
     3481 tctcaacccg ctcgcttcgt tccggtgggc ccggggcatg accgtcgtcg ccgccctgat
     3541 ggcggtcttc ttcatcatgc aacttgtcgg acaggtgccg gccgcgcttt gggtcatttt
     3601 cggcgaggat cgctttcact gggacgcgac cacgatcggc atttcgcttg ccgcatttgg
     3661 cattctgcat tcactcgccc aggcaatgat caccggccct gtagccgccc ggctcggcga
     3721 aaggcgggca ctcatgctcg gaatgattgc cgacggcaca ggctacatcc tgcttgcctt
     3781 cgcgacacgg ggatggatgg cgttcccgat catggtcctg cttgcttcgg gtggcatcgg
     3841 aatgccggcg ctgcaagcaa tgttgtccag gcaggtggat gaggaacgtc aggggcagct
     3901 gcaaggctca ctggcggcgc tcaccagcct gacctcgatc gtcggacccc tcctcttcac
     3961 ggcgatctat gcggcttcta taacaacgtg gaacgggtgg gcatggattg caggcgctgc
     4021 cctctacttg ctctgcctgc cggcgctgcg tcgcgggctt tggagcggcg cagggcaacg
     4081 agccgatcgc tgatcgtgga aacgataggc ctatgccatg cgggtcaagg cgacttccgg
     4141 caagctatac gcgccctagg agtgcggttg gaacgttggc ccagccagat actcccgatc
     4201 acgagcagga cgccgatgat ttgaagcgca ctcagcgtct gatcttaatt aaaacgtgag
     4261 accgaaagtg aaacgtgatt tcatgcgtca ttttgaacat tttgtaaatc ttatttaata
     4321 atgtgtgcgg caattcacat ttaatttatg aatgttttct taacatcgcg gcaactcaag
     4381 aaacggcagg ttcggatctt agctactaga gaaagaggag aaatactaga tgcgtaaagg
     4441 cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg
     4501 tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct
     4561 gaagttcatc tgtactactg gtaaactgcc ggttccttgg ccgactctgg taacgacgct
     4621 gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt
     4681 caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg
     4741 cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga
     4801 gctgaaaggc attgacttta aagaggacgg caatatcctg ggccataagc tggaatacaa
     4861 ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa
     4921 ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca
     4981 aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca
     5041 aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt
     5101 aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgaccag gcatcaaata
     5161 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     5221 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggtct
     5281 ctgagtcata tgcccactaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     5341 cttctgctta atttgatggt tatcagaaga gtgtaagcgg gttgactacc gaggaactga
     5401 ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac
     5461 caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac
     5521 gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc
     5581 ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt
     5641 ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat
     5701 caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag
     5761 cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag
     5821 gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc
     5881 ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta
     5941 ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta
     6001 ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg
     6061 gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg
     6121 catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg
     6181 cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg ccaggtcctc
     6241 gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt
     6301 cgccaaacct gccgcctcct gttcgagacg acgcgaacgc tccacggcgg ccgatggcgc
     6361 gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg
     6421 gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc
     6481 gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt
     6541 ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc acgccggggc
     6601 aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat aatccacctt
     6661 atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact tggtattccg
     6721 aatcttgccc tgcacgaata ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg
     6781 gacgtgcttg gcaatcacgc gcaccccccg gccgttttag cggctaaaaa agtcatggct
     6841 ctgccctcgg gcggaccacg cccatcatga ccttgccaag ctcgtcctgc ttctcttcga
     6901 tcttcgccag cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg
     6961 tgagccagag tttcagcagg ccgcccaggc ggcccaggtc gccattgatg cgggccagct
     7021 cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca
     7081 ggtaggccta caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt
     7141 ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag
     7201 ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat
     7261 tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg
     7321 gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta
     7381 cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa
     7441 tcgctataat gaccccgaag cagggttatg cagcggaaaa gatccgtc
//
