LOCUS       sequence-463776-        2940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VR_I11 (Chloramphenicol R) containing the insert I11 to be 
            assembled in POC1520 (pMOBK360_VR) using the site-selective 
            methylation protection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2940)
  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 2940)
  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: POC1545
FEATURES             Location/Qualifiers
     source          1..2940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1327..1429
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1430..2089
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1496..1515)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2115..2660
                     /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     2561..2580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2777..2794
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgacta ggtctcatga aaaagaccca aggcagggat
       61 gggagccctt cttgcagcag gagtggatgc aggacctgcc tggaagcaag agaaggacga
      121 gggaccctgg ctgggcccgt atttttagta gagacagttt ctccgtattg gtcaggctgg
      181 tctcgaactc ctgacctcag gtgattgcct cacagttcaa gtcaggcttg gggaaagtag
      241 tgagaccctg tctccagcca aaaaaaaaaa attaaattaa aatatactgt actataatag
      301 tgttaattgt aagcacagtg tagtgcaacc gatctttaga atatttcacc tactggccgg
      361 tgctgggttt cagcatcctg caatttcaga actactgtga atacaaaaga aagaaaggtc
      421 ctgcccagga tgggagtcac tctaattgcc attgagaaaa atatgcctag gccagaagat
      481 tggtgatggt gggaatagca gggacatagt ttagacaagg tcattgagca agacgtttct
      541 gagaacttga ccactggtta tctgagaaga gacctgaatc gtgtgaggag tgagtcatgt
      601 gaatccctgg ggagcaggta catgtgggag tcagagcatg ggtaggtggg atgttcaaag
      661 tgaagttgac atctacaata agtagtaagt gtagacctca gaaatttaac aagattctac
      721 tttttccccc aaaaaaatgt cctcacaatt tcctgctgaa aggagagggt tcattcaggg
      781 gaccacatga tgccgttctt ctgctgcagt tctaaattaa gttgtccttc cctctaacta
      841 caggtctcac aaacccttgc agccagcttc aatgatttta gcttgactgt ttttgacttg
      901 ggtattttgc caaaatggta tgtttttcta tagaatttga cgttgtggag accctgcttt
      961 tctgccaggt ccccagaggg gatacagtgc tttgtacggc aatgacattc ccctgagcta
     1021 tatccctggt tactacagca tttcctgatc ttgcctaaca ggattacttc cctgaactgt
     1081 aaaattcctg cagcattgta tacagagaaa gaataggaga catggcaatc atggacaaga
     1141 aaggaggaaa tcgtgtgaga ctgagaccta gtcgtgagag gtactgcacc acgaagtgtg
     1201 cggccgcctt cagcggccgc acgaataaat acctgtgacg gaagatcact tcgcagaata
     1261 aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg
     1321 agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1381 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1441 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1501 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1561 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1621 ccgcctgatg aatgctcatc cggagttccg tatggcaatg aaagacggtg agctggtgat
     1681 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1741 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1801 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1861 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     1921 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     1981 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2041 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaag agcctgttta
     2101 ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa
     2161 aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga
     2221 ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg
     2281 gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc
     2341 ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2401 ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact
     2461 gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc
     2521 gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt
     2581 atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag
     2641 gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat
     2701 cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag
     2761 tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat
     2821 attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca
     2881 ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgcgca agcggccgca
//
