LOCUS       sequence-463777-        2962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VL_I10 (Chloramphenicol R) containing the insert I10 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 2962)
  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 2962)
  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: POC1544
FEATURES             Location/Qualifiers
     source          1..2962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1349..1451
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1452..2111
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1518..1537)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2137..2682
                     /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     2583..2602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2799..2816
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcgttc acaggtgtca cccttacact
       61 ctctgccatg accaccttac gtgggtgggg ctgaggttgc cttgaccaag aagtgcatca
      121 gtttaaaatg taataaaata ggccgggtat ggtggctcat gcttgtaatc ccagcacttt
      181 gggaggctga ggcaggcaga tcacctgagg tcaggagttt gagaccagcc tggccaacat
      241 ggcaaaatcc tgtctctatg aaaaatgcaa aaattagccg ggtatggtgg tgcatgtgcc
      301 tcagctcctg agtagctggg actacaggca cacgccacca ggcccagcta attttttgta
      361 tttttagtag agacagggtt tcaccatgtt gatcaggatg gtctcgatct cttgacctga
      421 ttttctccat gtggagtagg ggagacccct tagcctgagg ctgcctctgc ccaccctctg
      481 cacctgggaa ctgccactgc cacagccacc atctccacac agaacctcct ggagaggaaa
      541 gggaaattgg ctcagggaga aagagtaggg cctcgaatgg tgcagaggac acgacaaccc
      601 caagaaagct cctggaggag gtgcatgtgc aaagtggaca gagatggcca tgtgtgctca
      661 ggaccctccc tgttacaagg ggtctccaaa gggctacaca ggcagggccc ccaggctggg
      721 cctcacagct gccttcatcc aggcccttag cattcagcac gtgattccca gctcacccca
      781 cctggatgcg ccctagtgag ttcaagagac acagagaccg ggatggggac agagcaggcg
      841 ccgcagccct ccctgctgcc cactcgcctc cctgtgtggg ctgagtgccg gctcacctcc
      901 ctgctgagtt ccactgaccc ccattcctca cccccagcct gagccagatc cagtgggaag
      961 agacaggtcc ctgctctctg cccctaactt ttctggaaaa ggcctctcca gtcactcttg
     1021 cccctgccga ctctcacctt ctttctggtc cttgatatga gccagggtca tcctgaggtc
     1081 ctttccgttc cctgtcaagt ccctggtctc tctgtcccat gtcttatttc ccagcacggg
     1141 ctctggggaa gaggaaatca caggtgaaac ttcttcctgg gagcgagacc aagtcgtgag
     1201 aggtactgca ccacgaagtg tgcggccgcc ttcagcggcc gcacgaataa atacctgtga
     1261 cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc gggaagccct
     1321 gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc aactttcacc
     1381 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     1441 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     1501 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     1561 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     1621 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggagttc cgtatggcaa
     1681 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1741 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1801 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     1861 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     1921 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     1981 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     2041 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     2101 gcggggcgta agagcctgtt taataagatg atcttcttga gatcgttttg gtctgcgcgt
     2161 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
     2221 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt
     2281 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
     2341 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
     2401 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
     2461 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
     2521 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
     2581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
     2641 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
     2701 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
     2761 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
     2821 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
     2881 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc
     2941 cgctagcgcg caagcggccg ca
//
