LOCUS       sequence-463779-        2710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VM_I3 (Chloramphenicol R) containing the insert I3 to be 
            assembled in POC1518 (pMOBK360_VL) using the site-selective 
            methylation protection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2710)
  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 2710)
  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: POC1537
FEATURES             Location/Qualifiers
     source          1..2710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1097..1199
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1200..1859
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1266..1285)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1885..2430
                     /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     2331..2350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2547..2564
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcgaac cctggaccag ttctaagggg
       61 tgtgatcaga ccacacccag agcagactgc acagttctgg tcccacgtct agaggacact
      121 cagacaagga acccccatat tccttaaatg taccttgagc acatgttctc agggtctcct
      181 gagggctgtg tcatgggctg tggtcactca ttgttggtct ctgtcaggaa gttgctatag
      241 ccatgtaaac caaaaaacgt ctgagacagg tctcagtcag tttagaagtt tatttcacca
      301 agctgaagat gcaccttgga aacagagaca caaatcattg tagcatttgg gggaaacatt
      361 caaaccatag cagttataaa ggatacaaat gagcagccag atgaagaggt gcacagtgag
      421 gcctggaagg gctctgagca caagagcctc tgtccccacg gagacaagtg cacaccaaca
      481 tgcccaacta atttttaaat attttgtagg aatgggtctc actattctgc ccagtctggt
      541 ctcaaactcc tggacttaaa caatgttcct accattgcct cccaaactga tggaattata
      601 agtgtgagcc actgtgcctg gccaatcaca gatctcctga tcctggcaca ttaccagaat
      661 cctctttggt cattaacaat tatcccagtg caatccataa acaggaggat atgcaggtgc
      721 attttctggt gtaaagaaat atgactcggt ggtccacatg gttgcagtac ctcccattca
      781 agttcttttg taacactcga gcatttgcat ctgagataaa gcagggacct cttttagaag
      841 cctgctgggg cctcccaatc tagaaataaa gaaaaacctt cagtcccttc aagagaaatt
      901 ccaggcacct agccagccct aaaaagtaag tcgagaccaa gtcgtgagag gtactgcacc
      961 acgaagtgtg cggccgcctt cagcggccgc acgaataaat acctgtgacg gaagatcact
     1021 tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt
     1081 ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     1141 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     1201 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     1261 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     1321 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     1381 acattcttgc ccgcctgatg aatgctcatc cggagttccg tatggcaatg aaagacggtg
     1441 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1501 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1561 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1621 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1681 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1741 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1801 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaag
     1861 agcctgttta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc
     1921 tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc
     1981 tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta
     2041 gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg
     2101 ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg
     2161 cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct
     2221 acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca
     2281 ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc
     2341 tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga
     2401 tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc
     2461 tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg
     2521 ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc
     2581 tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac
     2641 ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgcgca
     2701 agcggccgca
//
