LOCUS       Exported                5707 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    UbcH5c WT pET28a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5707)
  AUTHORS   Brzovic PS, Lissounov A, Christensen DE, Hoyt DW, Klevit RE
  TITLE     A UbcH5/ubiquitin noncovalent complex is required for processive 
            BRCA1-directed ubiquitination.
  JOURNAL   Mol Cell. 2006 Mar 17. 21(6):873-80.
  PUBMED    16543155
REFERENCE   2  (bases 1 to 5707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5707
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     89..111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(105..296)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1105..2187)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2149..2168
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2188..2265)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2471..2490
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2574..2593
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2574..2592
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2593..2617
                     /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)."
     RBS             2632..2654
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3145..3162
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3215..3233)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3229..3276
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3313..3768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3400..3419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3610..3631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3860..4675)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4583..4602
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4667..4686
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4797..5385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5286..5305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5539..5556
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5571..3
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5636..5655)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
       61 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
      121 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
      181 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
      241 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
      301 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
      361 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
      421 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
      481 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
      541 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
      601 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
      661 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
      721 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
      781 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
      841 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
      901 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
      961 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     1021 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     1081 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1141 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     1201 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     1261 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     1321 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     1381 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     1441 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     1501 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     1561 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     1621 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     1681 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     1741 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     1801 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     1861 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     1921 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     1981 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     2041 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     2101 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     2161 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     2221 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     2281 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     2341 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     2401 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     2461 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     2521 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     2581 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     2641 ctttaagaag gagatatacc atggcgctga aacggattaa taaggaactt agtgatttgg
     2701 cccgtgaccc tccagcacaa tgttctgcag gtccagttgg ggatgatatg tttcattggc
     2761 aagccacaat tatgggacct aatgacagcc catatcaagg cggtgtattc tttttgacaa
     2821 ttcattttcc tacagactac cccttcaaac cacctaaggt tgcatttaca acaagaattt
     2881 atcatccaaa tattaacagt aatggcagca tttgtctcga tattctaaga tcacagtggt
     2941 cgcctgcttt aacaatttct aaagttcttt tatccatttg ttcactgcta tgtgatccaa
     3001 acccagatga ccccctagtg ccagagattg cacggatcta taaaacagac agagataagt
     3061 acaacagaat atctcgggaa tggactcaga agtatgccat gtgatgctac cttaaagtca
     3121 gaataacctg cattatagct cgagcaccac caccaccacc actgagatcc ggctgctaac
     3181 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     3241 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     3301 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3421 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3601 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3721 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     3781 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3841 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     3901 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     3961 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     4021 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     4081 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     4141 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     4201 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     4261 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     4321 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     4381 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     4441 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     4501 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     4561 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     4621 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     4681 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     4741 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4801 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4861 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4921 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4981 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5041 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5101 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5161 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5221 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5281 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5341 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5401 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5461 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5521 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     5581 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     5641 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     5701 catggct
//
