LOCUS       sequence-210664-        4443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synthetic biology; gateway destination vector assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  AUTHORS   Gantner J, Ordon J, Ilse T, Kretschmer C, Gruetzner R, Lofke C, 
            Dagdas Y, Burstenbinder K, Marillonnet S, Stuttmann J
  TITLE     Peripheral infrastructure vectors and an extended set of plant parts
            for the Modular Cloning system.
  JOURNAL   PLoS One. 2018 May 30;13(5):e0197185. doi: 
            10.1371/journal.pone.0197185. eCollection 2018.
  PUBMED    29847550
REFERENCE   2  (bases 1 to 4443)
  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: "PLoS One.";
            date: "2018-05-30"; volume: "13(5):e0197185. doi"; pages: " 
            10.1371/journal.pone.0197185. eCollection 2018"
COMMENT     Sequence Label: pJOG562
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(63..651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(143..162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(798..1607)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1521..1540
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1599..1618
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1720..1736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1741..1760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1741..1759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1817..1838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1853..1858,1859..1876,1877..1883)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1891..1907
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1896..1918
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1915..1931
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2003..2127
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2168..2473)
                     /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(2270..2289)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2815..3474)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3389..3408
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3475..3577)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3751..3875)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3926..3943)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3926..3942)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3935..3957)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4151..4170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4335..4352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
       61 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      481 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      661 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat
      721 tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac
      781 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
      841 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
      901 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
      961 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     1021 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     1081 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     1141 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     1201 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1261 tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg
     1321 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1381 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1441 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     1501 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1561 ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcataac accccttgta
     1621 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     1681 atgtaacatc agagattttg agacacgggc cagagctgcc aggaaacagc tatgaccatg
     1741 taatacgact cactataggg gatagctggc acgacaggtt tcccgactgg aaagcgggca
     1801 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     1861 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     1921 acagctatga ccatgattac gccaagcttg catgcaggcc tctgcagtcg acgggcccgg
     1981 gatccgattt tggtctcacg aaaccacttt gtacaagaaa gctgaacgag aaacgtaaaa
     2041 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     2101 taaaacacaa catatccagt cactatggtc gacctgcaga ctggctgtgt ataagggagc
     2161 ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt
     2221 ggctgagatc agccacttct tccccgataa cggaaaccgg cacactggcc atatcggtgg
     2281 tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt
     2341 tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa
     2401 taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa
     2461 ccttaaactg catttcacca gcccctgttc tcgtcagcaa aagagccgtt catttcaata
     2521 aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga
     2581 cgacgggctt cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg
     2641 ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat
     2701 acctcttttt gacatacttc gggtatacat atcagtatat attcttatac cgcaaagatc
     2761 agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatccac gcgtttacgc
     2821 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     2881 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     2941 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     3001 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     3061 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     3121 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     3181 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     3241 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     3301 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     3361 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     3421 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     3481 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     3541 attatggtga aagttggaac ctcttgcgtg ccgatcaacg tctcattttc gccaaaagtt
     3601 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
     3661 ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttagcggc
     3721 cgctaagttg gcagcatcac ccgcggccgc catagtgact ggatatgttg tgttttacag
     3781 tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt
     3841 tacgtttctc gttcagcttt tttgtacaaa cttgtctcct gagaccaaaa tctagatgca
     3901 ttcgcgaggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3961 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4021 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4081 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4141 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4201 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4261 tgaccgtctc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4321 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4381 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4441 aag
//
