LOCUS       sequence-463784-        2535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VL_I8 (Chloramphenicol R) containing the insert I8 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 2535)
  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 2535)
  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: POC1542
FEATURES             Location/Qualifiers
     source          1..2535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        922..1024
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1025..1684
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1091..1110)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1710..2255
                     /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     2156..2175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2372..2389
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcgatt ctgctttcct gtctgccaag
       61 ccattccagg caatcccttc atttggtaaa catttatcaa atacctactg tgtgcagggc
      121 attgttttaa gaggagctgg gactgagata agaggaaata aaccctcctt gccctcaagg
      181 tttatttcga taaaggttaa aaaattccat tagtttttaa aaagtttttc attttgaatt
      241 ttttgggggt ttgtttggtc tccttaggaa ttatctttta tttcttgata tagttcaaaa
      301 ttcaaaattc aaaatgttca aaaggtaaaa tgtctctctc ttaccctgtc tcatccccct
      361 tttcacacaa atttcagggc acggtgcatt tattgaatca gagacatcac taagggatgg
      421 aagcactcag acaaggggac aaacaagagc ccattgagct acacattaca gttgcccaca
      481 ggactgtatg ggcctagaga caagcaggga agaagagcac tcccctcctc tccaggcagg
      541 agtgatggac cctcatcccc aggacgaggc agggctgctg atacacagta agggcaggag
      601 gagtttgtgt ggaacccaga gattcacttg ggggcctcct tgtttccctt gtcccattgt
      661 aaatgtgaac agaatcgtcc agcaacccag tctgacagcg tttgatttcc aaaggcccag
      721 acccctcagg aggaagattg gagccgcatt tcggagcgag accaagtcgt gagaggtact
      781 gcaccacgaa gtgtgcggcc gccttcagcg gccgcacgaa taaatacctg tgacggaaga
      841 tcacttcgca gaataaataa atcctggtgt ccctgttgat accgggaagc cctgggccaa
      901 cttttggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc accataatga
      961 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     1021 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     1081 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     1141 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1201 tattcacatt cttgcccgcc tgatgaatgc tcatccggag ttccgtatgg caatgaaaga
     1261 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1321 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1381 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     1441 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     1501 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     1561 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     1621 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     1681 gtaagagcct gtttaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct
     1741 tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc
     1801 aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca
     1861 gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc
     1921 tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga
     1981 taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac
     2041 tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa
     2101 tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa
     2161 acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt
     2221 cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac
     2281 ttccctgtta agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat
     2341 ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata
     2401 tatcctgtat cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga
     2461 agcacttcac tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc
     2521 gcgcaagcgg ccgca
//
