LOCUS       Exported                4200 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  dummy module (ccdB) for 2nd generation recipient assembly.
ACCESSION   .
VERSION     .
KEYWORDS    pJOG294
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4200)
  AUTHORS   Ordon J, Gantner J, Kemna J, Schwalgun L, Reschke M, Streubel J, 
            Boch J, Stuttmann J
  TITLE     Generation of chromosomal deletions in dicotyledonous plants 
            employing a user-friendly genome editing toolkit.
  JOURNAL   Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319.
  PUBMED    27579989
REFERENCE   2  (bases 1 to 4200)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4200
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        461..563
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             564..1223
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(630..649)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1565..1870
                     /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     1749..1768
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(1979..1995)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1979..1995)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1992..2014)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2003..2019
                     /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)."
     promoter        complement(2027..2057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2072..2093
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2210..2227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2381..2969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2461..2480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3140..4000)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3763..3782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4001..4105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4173..4191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga ttttgaagac aattgcagag accgagacgt tgatcggcac gtaagaggtt
      481 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
      541 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
      601 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
      661 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
      721 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
      781 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
      841 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
      901 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
      961 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     1021 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     1081 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     1141 atgccgtctg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     1201 atgagtggca gggcggggcg taatcgcgtg gatccggctt actaaaagcc agataacagt
     1261 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     1321 gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     1381 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     1441 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     1501 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg
     1561 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     1621 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     1681 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     1741 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtttccgtta tcggggaaga
     1801 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     1861 gggaatataa atgtcaggct cccttataca cagccagtcg gtctcagctt ttgtcttcaa
     1921 aatcggatcc cgggcccgtc gactgcagag gcctgcatgc aagcttggcg taatcatggt
     1981 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2041 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2101 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2161 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2221 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2281 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2341 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2401 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2461 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2521 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2581 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2641 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2701 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2761 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2821 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2881 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2941 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3001 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3061 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3121 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3181 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3241 agggcttacc atctggcccc agtgctgcaa tgataccgcg agagccacgc tcaccggctc
     3301 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3361 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3421 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3481 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3541 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3601 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3661 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3721 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3781 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3841 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3901 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3961 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4021 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4081 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4141 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
