LOCUS       sequence-463786-        2919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VL_I5 (Chloramphenicol R) containing the insert I5 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 2919)
  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 2919)
  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: POC1539
FEATURES             Location/Qualifiers
     source          1..2919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1306..1408
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1409..2068
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1475..1494)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2094..2639
                     /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     2540..2559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2756..2773
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcgaag ctgaaaacaa tggaaccatg
       61 ccttcagaat tccagaaaga aagttatttt caacgaataa atctatattc agccaaataa
      121 tcaagggtga aggtaaaata atacattttt agagactcag gggttacctt tatttttata
      181 tcttttagag acgaggttct cacaatgttg ttcaagctgg tctccaactc ctgatctcaa
      241 gcaatcctct tgccttagcg tcctgaattg ctgggattac aggggtgagc tgtcatgccc
      301 agcttgtaat gtactgacac cttcaagatt tccttctcca agtgtggcat acatgaccct
      361 ttgccaattt ttgatagggc tgtttttttt cttgtaaatt tgtttaagtt ccttgtaaac
      421 atacatgtga gctctcatca ttcttgttta aacacctaag aggcatccaa atcagtgcaa
      481 catggcaaga aaatgaaata agaaaccaat agaaggacca ggcatggtgg ctcatgcctg
      541 taatccctgc attttgggag gttgaggtgg aaggatcact tgagttcagg agtttgagac
      601 cagcctgata gtgagacctc atctctacca aataaaaata attttaaaag aaaaaagatc
      661 aatagatagg aaaggaagaa acaaaagaat ggcatcatct cacacatcgt tatgctaaag
      721 gagagaccaa acgaaggact acataacata cgactgcacg tccattacat tctagaaatt
      781 tcagtattgc agtgacagaa ggggagagga gggtgtgagg gaagaggaca gagagaaggg
      841 ctggggaagc atacaacaat aatagcacaa tggtaggtta caagaaacta cactgttaga
      901 agtgtccttt attttgctgg atgcagctta atataacctg aacttcactg tgaaaagtca
      961 aggaatcggg tttccattct ttgctattgt gtaaagaaat atagccaaaa gccactagga
     1021 gaattaaaac cataaactaa aaaatgttta tttgacacat aagaaagtag taaaggagga
     1081 acagaaacaa aaagatatga gacaaattga aaacgtatag caaaatggta gaccaaggag
     1141 cgagaccaag tcgtgagagg tactgcacca cgaagtgtgc ggccgccttc agcggccgca
     1201 cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg gtgtccctgt
     1261 tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag
     1321 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
     1381 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     1441 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     1501 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1561 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     1621 ggagttccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     1681 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     1741 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1801 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     1861 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     1921 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     1981 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     2041 ctgcgatgag tggcagggcg gggcgtaaga gcctgtttaa taagatgatc ttcttgagat
     2101 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     2161 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     2221 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     2281 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     2341 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     2401 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     2461 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     2521 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2581 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     2641 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     2701 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     2761 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     2821 agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag
     2881 taagccagta tacactccgc tagcgcgcaa gcggccgca
//
