LOCUS       sequence-463778-        2687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VR_I4 (Chloramphenicol R) containing the insert I4 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 2687)
  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 2687)
  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: POC1538
FEATURES             Location/Qualifiers
     source          1..2687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1074..1176
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1177..1836
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1243..1262)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1862..2407
                     /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     2308..2327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2524..2541
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgacta ggtctcagtc tcaccaaaac ccaaccattg
       61 caagtgaaga aatgacatta gtgtgctacc tactgtatta acaaacaaaa aggaaactgg
      121 tctctatgag aatccctatg cgtgaggagt atttaaatta ggagggtcgg gcgggtctca
      181 gcccctcctc acccccaggc tcccactcca tgaggtattt ctacacctcc gtgttatata
      241 aatttaattt atatttaaat ttaaagtata tatactagta aaatttatat acatatttta
      301 agatatatca aatattaatt ccgcggggcc ggggccaggg tctcacaccc tccagagcat
      361 gtacggctgc gacgtagtat taatatatat ctagtaaatt gggcatgacc agtacgccta
      421 cgacggcaag gattacatcg ccctgaacga ggacctgcgc tcctggatca gagactcgaa
      481 ctttccaatc caaatggatg gggcctgtgc ccatttggtg ttgagtccat catgggcttt
      541 atgtggttag tcctcagctg ggtcaccttc actgctccat tgtccttgtc ccttcagtgg
      601 aaacttgtcc agtgggagct gtgaccacag aggctcacac atcgcccagg gcggcccctg
      661 cacacggggg tctctgtgca ttctgagaca aattttcaga gccattcacc tcctgctctg
      721 cttctagagc tccttttctg ctctgctctt ctgccctctc tccctgccct ggttctagtg
      781 gagctgctgt ggctggagaa atagatttag gaggtcacac catatatatg agacttaaaa
      841 cctcaggcat ggatgaagca ccaagggagt gactgactat ggaaaagaat gagcacaagg
      901 actgaacctg agacctagtc gtgagaggta ctgcaccacg aagtgtgcgg ccgccttcag
      961 cggccgcacg aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt
     1021 gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg
     1081 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     1141 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     1201 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     1261 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     1321 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     1381 gctcatccgg agttccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     1441 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     1501 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     1561 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     1621 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     1681 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     1741 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     1801 caacagtact gcgatgagtg gcagggcggg gcgtaagagc ctgtttaata agatgatctt
     1861 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     1921 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     1981 gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca
     2041 agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt
     2101 tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg
     2161 ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt
     2221 ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac
     2281 aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2341 gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct
     2401 atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct
     2461 tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag
     2521 cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc
     2581 accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc
     2641 caacatagta agccagtata cactccgcta gcgcgcaagc ggccgca
//
