LOCUS       sequence-390531-        3471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 Module 4 destination vector (reverse orientation MoClo 
            insert).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3471)
  AUTHORS   Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R,
            Gamo ME, Starker CG, Zhang F, Voytas DF
  TITLE     An extensible vector toolkit and parts library for advanced 
            engineering of plant genomes.
  JOURNAL   Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312.
  PUBMED    36896468
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: "Plant 
            Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312."
COMMENT     Sequence Label: pJMC-4-Reverse
FEATURES             Location/Qualifiers
     source          1..3471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(86..91,92..109,110..116)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             170..850
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1170..1475
                     /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     1354..1373
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        1553..1657
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2725..3313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3214..3233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3467..3471,1..13)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcagtgagc gaggaagcgg aagagcgccc acctgctttt cccaggcgcg ccagcgtgag
       61 accggccgca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
      121 gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
      181 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
      241 atttcagtca gtcgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
      301 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
      361 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
      421 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
      481 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
      541 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
      601 cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
      661 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
      721 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
      781 gcttaatgaa ttacaacagt actgcgatga gtggcaggcg gggcgtaatc tagaggatcc
      841 ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga
      901 atatatactg atatgtatac ccgaagtatg tcaaaaggag gtatgctatg aagcagcgta
      961 ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat
     1021 ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg
     1081 gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc
     1141 ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag
     1201 agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac
     1261 ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt
     1321 acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg
     1381 tgccggtttc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca
     1441 aaaacgccat taacctgatg ttctggggaa tataaggtct ctctccgagc tccctctttt
     1501 gcaggtgtga tgaaaagcct tagggtcagg tggcactttt cggggaaatg tgcgatgaac
     1561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctaatga gacaataacc
     1621 ctgataaatg cttcaattat attgaaaaag gaagagtatg agtatccaac atttccgtgt
     1681 cgcccttatt cccttttttg cggccttctg ccttcctgtt tttgctcacc cagaaacgct
     1741 ggtgaaagta aaagatgctg aagatcagtt gggtgcccga gtgggttaca ttgaactgga
     1801 tctcaacagc ggcaaaatct tagagagttt tcgccccgaa gaacgcttcc caatgatgag
     1861 cactttcaaa gttctgctat gtggcgcggt gttatcccgt attgatgctg ggcaagagca
     1921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga
     1981 aaagcatctt acggatggca tgacagtaag agaattatgt agtgctgcca taaccatgag
     2041 tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc
     2101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaactct
     2221 ccgcaaacta ttaactggcg aactacttac tcttgcttca cgccagcaac tcattgactg
     2281 gatggaggcg gacaaagttg caggaccact tctgcgctcg gcacttccgg ctggctggtt
     2341 tattgctgat aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg
     2401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2521 gtcagacgaa gggaaataaa tagtagcccg cctgatgtgc gggctttttt ttggtggtgg
     2581 ttaccagtgt cgcttagcta ccgtaggtcc tcttaggcgg tccgattagc gacatagtca
     2641 tcaatgcctt gacggcaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctactaac
     2821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3001 ctcaagacga tagttacctg ataaggcgca gcggtcgggc tgaacggggg gttcgtgctc
     3061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3421 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg a
//
