LOCUS       sequence-463785-        2611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VR_I9 (Chloramphenicol R) containing the insert I9 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 2611)
  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 2611)
  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: POC1543
FEATURES             Location/Qualifiers
     source          1..2611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        998..1100
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1101..1760
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1167..1186)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1786..2331
                     /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     2232..2251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2448..2465
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgacta ggtctcagaa tgagctgtac tctcagctac
       61 tcgaagctgg tccctgcttt aggctgcgct ccctttctgg gccccaggtc ccgccttcta
      121 aatctcccca ggtctccagc ccactggaat tttctcttcc aagcgtggcc ccgccctctc
      181 cgctcgtgat tggccctaag ttccgggccc cagtttcatt ggtcctccgc cacacttcag
      241 ccctgcttcc ccgtcacttt tcagtcctcc tcgggatcgc gcatcacctg cactttctgg
      301 tctcctcctg ctctttctct cctcgcgtct cctccgcttc ctctcacttt tcggacaaac
      361 cagtccttct gaggcccatg ggttcccggg ctgcctccgg ggctgctcct gtgaatggca
      421 ttcgagtgcc cttgttcgag ctccttatat gtttcagtta ctgatccttt gtcagatgaa
      481 tagtttgaaa atattttctc ccattcttgg atggtctctt cattttgttt attgtttcct
      541 ttgctgtgca gaagcctttt tacttgatat gatcccattt atgcaatttt actttggtta
      601 cctcccttgc ccctgcacac ccagtccttg cacctggcct acaagtttgc catcttcatt
      661 cccccttctt ctgttcatca gccccctcct ctatcctccc accctcacag ttttctttgt
      721 atatgaaatc ctcgttcttg tccctttgcc cgtgtgcatt tcctgcccca ggaaggttgg
      781 gacagcagac ctgtgtgtta aacatcaatg tgaagttact tccaggtgag actgagacct
      841 agtcgtgaga ggtactgcac cacgaagtgt gcggccgcct tcagcggccg cacgaataaa
      901 tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg
      961 ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta agaggttcca
     1021 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     1081 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     1141 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     1201 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     1261 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggagttcc
     1321 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     1381 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     1441 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     1501 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     1561 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     1621 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     1681 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     1741 agtggcaggg cggggcgtaa gagcctgttt aataagatga tcttcttgag atcgttttgg
     1801 tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa
     1861 ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc
     1921 aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa
     1981 atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa
     2041 gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt
     2101 ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc
     2161 ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg
     2221 gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga
     2281 tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt
     2341 gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc
     2401 cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga
     2461 gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt
     2521 ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag
     2581 tatacactcc gctagcgcgc aagcggccgc a
//
