LOCUS       sequence-403362-        6314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNA donor for transposition, shuttle vector for Corynebacterium 
            glutamicum / E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6314)
  AUTHORS   Yang S, Zhu J, Zhou X, Zhang J, Li Q, Bian F, Zhu J, Yan T, Wang X, 
            Zhang Y, Yang J, Jiang Y, Yang S
  TITLE     RNA-Guided DNA Transposition in Corynebacterium glutamicum and 
            Bacillus subtilis.
  JOURNAL   ACS Synth Biol. 2023 Jul 21;12(7):2198-2202. doi: 
            10.1021/acssynbio.3c00193. Epub 2023 Jun 22.
  PUBMED    37348112
REFERENCE   2  (bases 1 to 6314)
  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.3c00193"; journalName: "ACS Synth Biol"; date: 
            "2023-07-21- 21"; volume: "12"; issue: "7"; pages: "2198-2202"
COMMENT     Sequence Label: pCgVchDonor
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1982..2427
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(2428..2514,2515..3849)
                     /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
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     rep_origin      3924..4512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4413..4432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4666..4683
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctagactcg aggagatttt tgagggggag ggagtcgagg aagagccaga gcagaaggcg
       61 ggggaaccgt tctctgccga cagcgtgagc ccccttaaaa atcaggccgg ggaggaaccg
      121 gggagggatc agagctagga gcgagacacc ctaaaggggg ggaaccgttt tctgctgacg
      181 gtgtttcgtt tattagtttt cagcccgtgg atagcggagg gtgagggcaa gtgagagcca
      241 gagcaaggac gggaccccta aaggggggaa ccgttttctg ctgacggtgt ttcgtttatt
      301 agttttcagc ccgtggacgg ccgcgtttag cttccattcc aagtgccttt ctgacttgtt
      361 ggatgcgcct ttcactgaca cctagttcgc ctgcaagctc acgagtcgag ggatcagcaa
      421 ccgattgaga acgggcatcc aggatcgcag ttttgacgcg aagttcgagc aactcgcctg
      481 tcatttctcg gcgtttgttt gcttccgcta atcgctgtcg cgtctcctgc gcatacttac
      541 tttctgggtc agcccatctg cgtgcattcg atgtagctgc gccccgtcgc cccatcgtcg
      601 ctagagcttt ccgccctcgg ctgctctgcg tttccacccg acgagcaggg acgactggct
      661 ggcctttagc cacgtagccg cgcacacgac gcgccatcgt caggcgatca cgcatggcgg
      721 gaagatccgg ctcccggccg tctgcaccga ccgcctgggc aacgttgtac gccacttcat
      781 acgcgtcgat gatcttggta tcttttaggc gctcaccagc agctttgagc tggtatccca
      841 cggtcaacgc gtggcgaaac gcggtctcgt cgcgcgctgc gcgctctgga tttgtccaga
      901 gcactcgcac gccgtcgatc aggtcgccgg acgcgtccag ggcgctcggc aggctcgcgt
      961 ccaaaatcgc tagcgccttg gcttctgcgg tggcgcgttg tgccgcttca atgcgggcgc
     1021 gtccgctgga aaagtcctgc tcaatgtact ttttcggctt ctgtgatccg gtcatcgttc
     1081 gagcaatctc cattaggtcg gccagccgat ccacacgatc atgctggcag tgccatttat
     1141 aggctgtcgg atcgtctgag acgtgcagcg gccaccggct cagcctatgc gaaaaagcct
     1201 ggtcagcgcc gaaaacacga gtcatttctt ccgtcgttgc agccagcagg cgcatatttg
     1261 ggctggtttt acctgctgcg gcatacaccg ggtcaatgag ccagatgagc tggcatttcc
     1321 cgctcagcgg attcacgccg atccaagccg gcgctttttc taggcgtgcc catttctcta
     1381 aaatcgcgta gacctgcggg tttacgtgct caatcttccc gccggcctgg tggctgggca
     1441 catcgatgtc aagcacgatc accgcggcat gttgcgcgtg cgtcagcgca acgtactggc
     1501 accgcgtcag cgcttttgag ccagcccggt agagctttgg ttgggtttcg ccggtatccg
     1561 ggtttttaat ccaggcgctc gcgaaatctc ttgtcttgct gccctggaag ctttcgcgtc
     1621 ccaggtgagc gagcagttcg cggcgatctt ctgccgtcca gccgcgtgag cgcagcgcta
     1681 gttcggggtg ggtgtcgaac agatcggcgg aaaatttcca cgcgctagct gtgactgtgt
     1741 cctgcggatc ggctagagtc atgtcttgag tgctttctcc cagctgatga ctgggggtta
     1801 gccgacgccc tgtgagttcc cgctcacggg gcgttcaact ttttcaggta tttgtgcagc
     1861 ttatcgtgtt ttcttcgtaa atgaacgctt aactaccttg ttaaacgtgg caaataggca
     1921 ggattgatgg ctttcataga aggcggcggt ggaatcgaaa tctcgatcct ttttaaccca
     1981 tcacatatac ctgccgttca ctattattta gtgaaatgag atattatgat attttctgaa
     2041 ttgtgattaa aaaggcaact ttatgcccat gcaacagaaa ctataaaaaa tacagagaat
     2101 gaaaagaaac agatagattt tttagttctt taggcccgta gtctgcaaat ccttttatga
     2161 ttttctatca aacaaaagag gaaaatagac cagttgcaat ccaaacgaga gtctaataga
     2221 atgaggtcga aaagtaaatc gcgcgggttt gttactgata aagcaggcaa gacctaaaat
     2281 gtgtaaaggg caaagtgtat actttggcgt caccccttac atattttagg tcttttttta
     2341 ttgtgcgtaa ctaacttgcc atcttcaaac aggagggctg gaagaagcag accgctaaca
     2401 cagtacataa aaaaggagac atgaacgatg aacatcaaaa agtttgcaaa acaagcaaca
     2461 gtattaacct ttactaccgc actgctggca ggaggcgcaa ctcaagcgtt tgcgaaagaa
     2521 acgaaccaaa agccatataa ggaaacatac ggcatttccc atattacacg ccatgatatg
     2581 ctgcaaatcc ctgaacagca aaaaaatgaa aaatatcaag tttctgaatt tgattcgtcc
     2641 acaattaaaa atatctcttc tgcaaaaggc ctggacgttt gggacagctg gccattacaa
     2701 aacgctgacg gcactgtcgc aaactatcac ggctaccaca tcgtctttgc attagccgga
     2761 gatcctaaaa atgcggatga cacatcgatt tacatgttct atcaaaaagt cggcgaaact
     2821 tctattgaca gctggaaaaa cgctggccgc gtctttaaag acagcgacaa attcgatgca
     2881 aatgattcta tcctaaaaga ccaaacacaa gaatggtcag gttcagccac atttacatct
     2941 gacggaaaaa tccgtttatt ctacactgat ttctccggta aacattacgg caaacaaaca
     3001 ctgacaactg cacaagttaa cgtatcagca tcagacagct ctttgaacat caacggtgta
     3061 gaggattata aatcaatctt tgacggtgac ggaaaaacgt atcaaaatgt acagcagttc
     3121 atcgatgaag gcaactacag ctcaggcgac aaccatacgc tgagagatcc tcactacgta
     3181 gaagataaag gccacaaata cttagtattt gaagcaaaca ctggaactga agatggctac
     3241 caaggcgaag aatctttatt taacaaagca tactatggca aaagcacatc attcttccgt
     3301 caagaaagtc aaaaacttct gcaaagcgat aaaaaacgca cggctgagtt agcaaacggc
     3361 gctctcggta tgattgagct aaacgatgat tacacactga aaaaagtgat gaaaccgctg
     3421 attgcatcta acacagtaac agatgaaatt gaacgcgcga acgtctttaa aatgaacggc
     3481 aaatggtacc tgttcactga ctcccgcgga tcaaaaatga cgattgacgg cattacgtct
     3541 aacgatattt acatgcttgg ttatgtttct aattctttaa ctggcccata caagccgctg
     3601 aacaaaactg gccttgtgtt aaaaatggat cttgatccta acgatgtaac ctttacttac
     3661 tcacacttcg ctgtacctca agcgaaagga aacaatgtcg tgattacaag ctatatgaca
     3721 aacagaggat tctacgcaga caaacaatca acgtttgcgc cgagcttcct gctgaacatc
     3781 aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca attaacagtt
     3841 aacaaataaa aacgcaaaag aaaatgccga tagttttcgt tccactgagc gtcagacccc
     3901 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4021 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4081 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4141 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4201 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4261 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4321 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4381 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4441 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4501 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4561 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4621 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4681 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4741 caccgcatat gaagcttaag agcgcaacgc aataagcttt gttgatacaa ccataaaatg
     4801 ataattacac ccataaattg ataattatca cacccataaa ttgatattgc ctcttcatgg
     4861 tctaaacttc agtaagttta cgacattttc atgaagaggt catttccggg gatcctgatg
     4921 gcaggttggg cgtcgcttgg tcggtcattt cgaagggcac caataactgc cttaaaaaaa
     4981 gatctgtaca tcgatatcat atgcaagggt ttattgtttt ctaaaatctg attaccaatt
     5041 agaatgaata tttcccaaat attaaataat aaaacaaaaa aattgaaaaa agtgtttcca
     5101 ccattttttc aattttttta taattttttt aatctgttat ttaaatagtt tatagttaaa
     5161 tttacatttt cattagtcca ttcaatattc tctccaagat aactacgaac tgctaacaaa
     5221 attctctccc tatgttctaa tggagaagat tcagccactg catttcccgc aatatctttt
     5281 ggtatgattt tacccgtgtc catagttaaa atcatacggc ataaagttaa tatagagttg
     5341 gtttcatcat cctgataatt atctattaat tcctctgacg aatccataat ggctcttctc
     5401 acatcagaaa atggaatatc aggtagtaat tcctctaagt cataatttcc gtatattctt
     5461 ttattttttc gttttgcttg gtaaagcatt atggttaaat ctgaatttaa ttccttctga
     5521 ggaatgtatc cttgttcata aagctcttgt aaccattctc cataaataaa ttcttgtttg
     5581 ggaggatgat tccacggtac catttcttgc tgaataataa ttgttaattc aatatatcgt
     5641 aagttgcttt tatctcctat tttttttgaa ataggtctaa ttttttgtat aagtatttct
     5701 ttactttgat ctgtcaatgg ttcagatacg acgactaaaa agtcaagatc actatttggt
     5761 tttagtccac tctcaactcc tgatccaaac atgtaagtac caataaggtt attttttaaa
     5821 tgtttccgaa gtattttttt cactttatta atttgttcgt atgtattcaa atatatcctc
     5881 ctcactattt tgattagtac ctattttata tccatagttg ttaattaaat aaacttaatt
     5941 tagtttattt atagatttca ttggcttcta aattttttat ctagataata attattttag
     6001 ttaattttat tctagattat atatgatatg atctttcatt tccataaaac taaagtaagt
     6061 gtaaacctat tcattgtttt aaaaatatct cttgccagtc acgttacgtt attagttcat
     6121 gaccaaaatc ccttaacgtg ctagagtcct tactgcagta gttttgctga aatactcgat
     6181 tcacaaaaat atcaacttat ggttgttttg tgagatatca atatatggtt gttttgtggt
     6241 taagttgctg attataaata attattaaat atcactttat ggttgcatca acaggtaccg
     6301 agctcgaatt cact
//
