LOCUS       Exported                3539 bp ds-DNA     circular SYN 04-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    LI D-E TurboID
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3539)
  AUTHORS   Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske 
            M
  TITLE     A modular plasmid assembly kit for multigene expression, gene 
            silencing and silencing rescue in plants.
  JOURNAL   PLoS One. 2014 Feb 13;9(2):e88218. doi: 
            10.1371/journal.pone.0088218. eCollection 2014.
  PUBMED    24551083
REFERENCE   2  (bases 1 to 3539)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3539
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     466..484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(522..544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     644..663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     857..879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     871..888
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     872..888
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             950..976
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             980..1006
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1010..1036
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1043..1999
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     primer_bind     complement(2063..2079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2063..2079)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2076..2098)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2087..2103
                     /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(2111..2141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2156..2177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2294..2311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2465..3053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2545..2564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3168..3196
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3385..3539,1..379)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cgacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
       61 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
      121 ggcttacgtt ctgcccaggt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
      181 agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
      241 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
      301 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
      361 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
      421 gttcggtaaa ttgtcacaac gccgctcatg acattaacct ataaaaatag gcgtatcacg
      481 aggccctttt gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
      541 ccggagaagg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      601 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      661 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      721 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      781 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      841 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
      901 ggtacctcgc gaatgcatct agatatcgga tccccgggtc tcaaaggcat acccgtatga
      961 tgttccggat tacgctggct atccctacga cgtgcccgac tatgccgggt acccctatga
     1021 cgtcccagac tacgcagcta gcaaagacaa tactgtgcct ctgaagctga tcgctctcct
     1081 ggctaatggc gagttccata gtggcgaaca gctgggagaa accctgggca tgtccagggc
     1141 cgctatcaac aagcacattc agactctgcg cgactggggc gtggacgtgt tcaccgtgcc
     1201 cggaaagggc tactctctgc ccgagcctat cccgctgctg aacgctaaac agattctggg
     1261 acagctggac ggcgggagcg tggcagtcct gcctgtggta gactccacca atcagtacct
     1321 gctggatcga atcggcgagc tgaagagtgg ggatgcttgc attgcagaat atcagcaggc
     1381 agggagagga agcagaggga ggaaatggtt ctctcctttt ggagctaacc tgtacctgag
     1441 tatgttttgg cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg tcatcggaat
     1501 tgtcatggca gaagcgctgc gaaagctggg agcagacaag gtgcgagtca aatggcccaa
     1561 tgacctgtat ctgcaggata gaaagctggc aggcatcctg gtggagctgg ccggaataac
     1621 aggcgatgct gcacagatcg tcattggcgc cgggattaac gtggctatga ggcgcgtgga
     1681 ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa gcagggatta acctggacag
     1741 gaatactctg gccgctacgc tgatccgaga gctgcgggca gccctggaac tgttcgagca
     1801 ggaaggcctg gctccatatc tgccacggtg ggagaagctg gataacttca tcaatagacc
     1861 cgtgaagctg atcattgggg acaaagagat tttcgggatt agccggggga ttgataaaca
     1921 gggagccctg ctgctggaac aggacggagt tatcaaaccc tggatgggcg gagaaatcag
     1981 tctgcggtct gccgaaaagt agaatctgag acctcgggcc cgtcgactgc agaggcctgc
     2041 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2101 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2161 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2221 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2281 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2341 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2401 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3121 tggtcatgag gcgcacaccg tggaaacgga tgaaggcacg aacccagttg acataagcct
     3181 gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     3241 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     3301 tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     3361 gtttgatgtt atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc
     3421 agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc
     3481 ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgg
//
