LOCUS       sequence-394770-        3471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level1 plasmid, allows transition into Cermak, et al., 2017 GG
            cloning system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3471)
  TITLE     Curtin lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3471)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJC226
FEATURES             Location/Qualifiers
     source          1..3471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(276..282,283..300,301..306))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     complement(379..396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(550..1138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(630..649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(2206..2310)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(2388..2693)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2490..2509)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(join(3013..3471,1..222))
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
ORIGIN
        1 acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
       61 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
      121 gttataggta cattgagcga ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
      181 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
      241 tgaaaatctc gacggatcct aactcaaaat ccacacatta tacgagccgg aagcataaag
      301 tgtaaagcct ggggtgccta atgcggccgg tctcactccg gcgcgccccg gaaaagcagg
      361 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      421 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      481 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      601 tcagaggtgg cgaaaccaga caggactata aagataccag gcgtttcccc ctggaagctc
      661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      781 cgttcgctcc aagctgggct gtgagcacga accccccgtt cagcccgacc gctgcgcctt
      841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      961 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1021 gccagttacc ttcggaaaaa gagttagtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1081 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1201 gattttgccg tcaaggcatt gatgactatg tcgctaatcg gaccgcctaa gaggacctac
     1261 ggtagctaag cgacactggt aaccaccacc aaaaaaaagc ccgcacatca ggcgggctac
     1321 tatttatttc ccttcgtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagagc cacgctcacc
     1501 ggctccagat ttatcagcaa taaaccagcc agccggaagt gccgagcgca gaagtggtcc
     1561 tgcaactttg tccgcctcca tccagtcaat gagttgctgg cgtgaagcaa gagtaagtag
     1621 ttcgccagtt aatagtttgc ggagagttgt tgccattgct acaggcatcg tggtgtcacg
     1681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccaatcg ttgtcagaag
     1801 taagttggcc gcagtgttat cactcatggt tatggcagca ctacataatt ctcttactgt
     1861 catgccatcc gtaagatgct tttctgtgac tggtgaatac taaaccaagt cattctgaga
     1921 atagtgtatg cggcgaccga gttgctcttg ccaagcatca atacgggata acaccgcgcc
     1981 acatagcaga actttgaaag tgctcatcat tgggaagcgt tcttcggggc gaaaactctc
     2041 taagattttg ccgctgttga gatccagttc aatgtaaccc actcgggcac ccaactgatc
     2101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcagaaggc
     2161 cgcaaaaaag ggaataaggg cgacacggaa atgttggata ctcatactct tcctttttca
     2221 atataattga agcatttatc agggttattg tctcattagc ggatacatat ttgaatgtat
     2281 ttagaaaaat aaacaaatag gggttcatcg cacatttccc cgaaaagtgc cacctgaccc
     2341 taaggctttt catcacacct gcaaaacact gagctcagcg agagacctta tattccccag
     2401 aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct
     2461 tccccgataa cggaaaccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca
     2521 tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag cagacgtgca
     2581 ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc
     2641 acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca
     2701 gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga cctcagccat
     2761 cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat
     2821 ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg
     2881 tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcctttt gacatacttc
     2941 gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact
     3001 gttatctggc ttttagtaag ccggatcctc tagattacgc cccgcctgcc actcatcgca
     3061 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     3121 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     3181 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     3241 cacccaggga ttggctgaaa cgaaaaacat attctcaata aaccctttag ggaaataggc
     3301 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     3361 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     3421 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca t
//
