LOCUS       sequence-463787-        2777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMOLC360_VM_I2 (Chloramphenicol R) containing the insert I2 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 2777)
  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 2777)
  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: POC1536
FEATURES             Location/Qualifiers
     source          1..2777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1164..1266
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1267..1926
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1333..1352)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1952..2497
                     /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     2398..2417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2614..2631
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactctgtgg tgcagtacct ctcacgactt ggtctcgaag tgagtgacct tataagcaag
       61 aaggaatgat agcttgaaac aatagccaag gaagttagag tcacaagatg tttcattctc
      121 tatagaaact aaaaataaca tcttaacata tgtccctgag ttgtttttca gaaacatgga
      181 ttccctccta atagatccac tgacagggag accttataaa ggggaaactg agaactgaac
      241 tctgaccgcc gttctttgtt ctaaatttct tcctgaggga cctggagaga gtcaagccca
      301 caggccaggc ctgaacattc ctctcctttg cttccttatc tatcacaaat cagagaatct
      361 ttaaatctac ctaagacctg taagctccaa aaaaaagccg caaaggatac aaataagaag
      421 tttacaaaag atatagattt aaaaaaaaaa acagaaattc tggaattaaa gaattcatga
      481 catgaaattc caaatacatt tgaaagcttc aacaacatac tagacacagc taattttttt
      541 tttttttttt gtatttttag tagagacggg atttcaccaa gttggccagg atggtctcaa
      601 tctcttgaca tcatgatcca cccacctcag cctcccaaag tgctgggatt acaggtgtga
      661 gccactgtgc ccagctctaa cagcagattt cttagcagaa acctgaagga aaagaactgc
      721 caagccaaga gtctacagga aatgctcaag aaagtcctaa acctggaatt gaaacgacat
      781 tttcaacatt tccacattat caccatgaaa acatacacaa gtataaaact cactggtaaa
      841 gcaatcacac aaaagaggaa gagaaaggtc tcaaatggtc ctactacaga aatccaccaa
      901 accataatga cacctggccc acccagggcg gaaaactgct taaagtcgtt cttaaaccac
      961 aaacaatagc atgagtgatc tgtgccttaa ggacatgccg agaccaagtc gtgagaggta
     1021 ctgcaccacg aagtgtgcgg ccgccttcag cggccgcacg aataaatacc tgtgacggaa
     1081 gatcacttcg cagaataaat aaatcctggt gtccctgttg ataccgggaa gccctgggcc
     1141 aacttttggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat
     1201 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     1261 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1321 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1381 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1441 tttattcaca ttcttgcccg cctgatgaat gctcatccgg agttccgtat ggcaatgaaa
     1501 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1561 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1621 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1681 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1741 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1801 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1861 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     1921 gcgtaagagc ctgtttaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct
     1981 cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta
     2041 ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt
     2101 cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg
     2161 gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg
     2221 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga
     2281 actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg
     2341 aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg
     2401 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat
     2461 ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc
     2521 acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc
     2581 atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa
     2641 tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat
     2701 gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta
     2761 gcgcgcaagc ggccgca
//
