LOCUS       sequence-463791-        3038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VM_I6 (Chloramphenicol R) containing the insert I6 to be 
            assembled in POC1519 (pMOBK360_VM) using the site-selective 
            methylation protection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3038)
  AUTHORS   Flores-Fernandez CN, Lin D, Robins K, O'Callaghan CA
  TITLE     UniClo: scarless hierarchical DNA assembly without sequence 
            constraint.
  JOURNAL   Nucleic Acids Res. 2025 Jun 20;53(12):gkaf548. doi: 
            10.1093/nar/gkaf548.
  PUBMED    40548934
REFERENCE   2  (bases 1 to 3038)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2025-06-20"; volume: "53(12):gkaf548. doi"; 
            pages: " 10.1093/nar/gkaf548"
COMMENT     Sequence Label: POC1540
FEATURES             Location/Qualifiers
     source          1..3038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1425..1527
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1528..2187
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1594..1613)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2213..2758
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2659..2678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2875..2892
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcgatg gcaaattatc ttaaaactgg
       61 ggaaaacttt ttcatgttca gattttaaaa aaaacctata atcactgaca tgtccaattt
      121 ctgatttttc aaataagata gatttcttgt ctgcaacaat aaaacagcct cccagaaaca
      181 tcactggatc ttactcagtc acaaaccacc ctccccaacc ctagagtagc cactcctgat
      241 ttctaacaca gataagttct gccagttcca gttgaatggc attatacagt caccatcatg
      301 ctcctgggtc tcatcatcct gctgtgtgtg gtcatagcct attcactccc atggctgtcc
      361 aatgctccac tttataaata gctcacattt cttgtccctt tgctgttgat agacaattgt
      421 gttatatcca gattttacaa atcgtgacac gcaaatgttg cagccttgtt tgtaacagtg
      481 aaaaattgaa aacaacctgg aagtccagtg atgggaaaat gaatatattt ctgtcttttg
      541 gggaacccaa agcaggttcc aagactgcaa tttcagtgaa agcagtcagt gaatggcaaa
      601 cacttaaagc aatttcatgt ttaagtatat aagctcttca tatctttttt ttttttttga
      661 cagagtttcg ctcttgttac cctgacggag tgcaatggca caacttcagc tcactgcaac
      721 ctctgacttc ccaggttcaa atgattctcc tgcctcaggc tcccatgtag ctgggactac
      781 aggcacgtgc cacgacgcct ggctaatttt gtgtttttgg tgaagacggg gtttcacaat
      841 gttggtcagg ctggtctcaa actcctgacc taaagtgatc cgcctgcctc ggcctcccaa
      901 agtgctggga ttataggctt gtttgagatg cctttgagaa atgaagaaag tagagagaga
      961 ataaaagaaa aactggccag gcacagtggc tcatgcctgt aatacagtgg ctcagcattt
     1021 tgggaggcag agacaggcag ataccttgag atcagcttgg gcaacatggt gaagccccat
     1081 ctctacaaaa aatacaaaag ttagctgggc attgtggcgc acacctgtag tcccatctac
     1141 tcagggagct gagatggaag gatcaattga gcctgcgagt tcgaggctgc agtgagttgt
     1201 gattgtgcca ctgcactcca gcctgggtga cagaagagac cttgtctcga aaaagaagcc
     1261 gagaccaagt cgtgagaggt actgcaccac gaagtgtgcg gccgccttca gcggccgcac
     1321 gaataaatac ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt
     1381 gataccggga agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga
     1441 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     1501 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1561 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1621 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1681 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1741 gagttccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1801 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1861 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1921 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1981 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     2041 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     2101 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     2161 tgcgatgagt ggcagggcgg ggcgtaagag cctgtttaat aagatgatct tcttgagatc
     2221 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     2281 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     2341 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     2401 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     2461 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     2521 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     2581 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     2641 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2701 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     2761 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     2821 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     2881 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
     2941 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
     3001 aagccagtat acactccgct agcgcgcaag cggccgca
//
