LOCUS       sequence-463790-        2600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VL_I1 (Chloramphenicol R) containing the insert I1 to be 
            assembled in POC1518 (pMOBK360_VL) using the site-selective 
            methylation protection approach.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2600)
  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 2600)
  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: POC1535
FEATURES             Location/Qualifiers
     source          1..2600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        987..1089
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1090..1749
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1156..1175)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1775..2320
                     /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     2221..2240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2437..2454
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcgctc catctataaa caggtgtttc
       61 ccagtctgaa agggtgtttc atttaaaaca ggtctaactt ctgtttgata actaattaaa
      121 tctaaacatc tgtgctctgg ggatggcttc atactttaaa attcttgaat cagtaacatc
      181 tggccagcta tttgtgacga agtgaagctt taacattcct tgtccaagtg ggtctcctcc
      241 atctaaacca gcatatttcc tcatttgttc cttaagcctg ttaaaaaatt ccacatattc
      301 tgggtgcagg gtaacgattc tctaattcct tttattatta tttcttgcag gtctctcata
      361 ttcctcctat gagctatgtt gttattatcc cactgggaat cttgagtagg aaatttctgt
      421 ttggctgcag ggacgttttg accaggagga tgataacctc ttgccattct tttaccttct
      481 gtctctatgg attgtggtgc tgtttacatt ccagattttt attaatatgt gaggctttgt
      541 gccgagacta gctcagttgg gaagacccta acccagtggc actagaggaa tcaaagacac
      601 acacacagaa atatagaggt gtgaagtggg aaatcaggga tctcacagcc ttcagagctg
      661 acagccttga acagagattt acccccatat ttattaactc aagccagtga taagcattgt
      721 ttctatagat tatagattaa ctaaaagtat tccttatggg aaacaaacgg atgggccaaa
      781 ataaagggat gggtttggct agttatctgc agcaggagca tcgagaccaa gtcgtgagag
      841 gtactgcacc acgaagtgtg cggccgcctt cagcggccgc acgaataaat acctgtgacg
      901 gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg
      961 gccaactttt ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat
     1021 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     1081 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1141 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1201 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1261 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggagttccg tatggcaatg
     1321 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1381 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1441 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1501 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     1561 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     1621 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1681 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1741 ggggcgtaag agcctgttta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa
     1801 tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag
     1861 ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc
     1921 tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca
     1981 gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta
     2041 ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag
     2101 cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag
     2161 cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg
     2221 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca
     2281 gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct
     2341 ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa
     2401 gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
     2461 gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca
     2521 catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg
     2581 ctagcgcgca agcggccgca
//
