LOCUS       sequence-464650-        3656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  XanthoMoClo Golden Gate Part Plasmid for cloning, contains 
            Marker_SacB_PartType4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3656)
  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 3656)
  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: pXTK014 [Marker_SacB_PartType4]
FEATURES             Location/Qualifiers
     source          1..3656
                     /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"
     promoter        620..1065
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1066..1152,1153..2487)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        2634..2725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2726..2794,2795..3586)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3590..3629
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
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 cgcgagtcgg tctcttggcc acatatacct gccgttcact attatttagt gaaatgagat
      661 attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc aacagaaact
      721 ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta ggcccgtagt
      781 ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca gttgcaatcc
      841 aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa
      901 gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca ccccttacat
      961 attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag gagggctgga
     1021 agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa catcaaaaag
     1081 tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg aggcgcaact
     1141 caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg catttcccat
     1201 attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt
     1261 cctgaattcg attcgtccac aattaaaaat atctcttctg caaaaggcct ggacgtttgg
     1321 gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg ctaccacatc
     1381 gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta catgttctat
     1441 caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt ctttaaagac
     1501 agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga atggtcaggt
     1561 tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt ctccggtaaa
     1621 cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc agacagctct
     1681 ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg aaaaacgtat
     1741 caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa ccatacgctg
     1801 agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga agcaaacact
     1861 ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata ctatggcaaa
     1921 agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg
     1981 gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta cacactgaaa
     2041 aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga acgcgcgaac
     2101 gtctttaaaa tgaacggcaa atggtacctg ttcactgact cccgcggatc aaaaatgacg
     2161 attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa ttctttaact
     2221 ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct tgatcctaac
     2281 gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg
     2341 attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac gtttgcgcca
     2401 agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa
     2461 caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgatg gccgcggcgt
     2521 tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattga
     2581 acgtgagacc agagtagccc gcctaatgag cgggcttttt tttaattccc ctatttgttt
     2641 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2701 tcaataatat tgaaaaagga agagtatgag cattcagcat tttcgtgtgg cgctgattcc
     2761 gttttttgcg gcgttttgcc tgccggtgtt tgcgcatccg gaaaccctgg tgaaagtgaa
     2821 agatgcggaa gatcaactgg gtgcgcgcgt gggctatatt gaactggatc tgaacagcgg
     2881 caaaattctg gaatcttttc gtccggaaga acgttttccg atgatgagca cctttaaagt
     2941 gctgctgtgc ggtgcggttc tgagccgtgt ggatgcgggc caggaacaac tgggccgtcg
     3001 tattcattat agccagaacg atctggtgga atatagcccg gtgaccgaaa aacatctgac
     3061 cgatggcatg accgtgcgtg aactgtgcag cgcggcgatt accatgagcg ataacaccgc
     3121 ggcgaacctg ctgctgacga ccattggcgg tccgaaagaa ctgaccgcgt ttctgcataa
     3181 catgggcgat catgtgaccc gtctggatcg ttgggaaccg gaactgaacg aagcgattcc
     3241 gaacgatgaa cgtgatacca ccatgccggc agcaatggcg accaccctgc gtaaactgct
     3301 gacgggtgag ctgctgaccc tggcaagccg ccagcaactg attgattgga tggaagcgga
     3361 taaagtggcg ggtccgctgc tgcgtagcgc gctgccggct ggctggttta ttgcggataa
     3421 aagcggtgcg ggcgaacgtg gcagccgtgg cattattgcg gcgctgggcc cggatggtaa
     3481 accgagccgt attgtggtga tttataccac cggcagccag gcgacgatgg atgaacgtaa
     3541 ccgtcagatt gcggaaattg gcgcgagcct gattaaacat tggtaaaccg atacaattaa
     3601 aggctccttt tggagccttt ttttttggac gacccttgtc agatcaaagg atcttc
//
