LOCUS       Exported                5239 bp ds-DNA     circular SYN 07-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  AUTHORS   Berta Canal
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Apr 7, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5239
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    115..192
                     /label=14His tag
     misc_feature    193..483
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
     misc_feature    484..1428
                     /label=Papain-Like protease (PLpro_nsp3)
                     /label=coPlPro-Nsp3
     terminator      1481..1575
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1636..2451
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2670..2756
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2850..3932)
                     /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"
     promoter        complement(3933..4010)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    4202..4342
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4528..5116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatgagc
      121 aagcatcacc atcattcagg ccatcaccat accggacacc accatcattc aggcagtcat
      181 caccataccg gatcggactc agaagtcaat caagaagcta agccagaggt caagccagaa
      241 gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga gatcttcttc
      301 aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa aagacagggt
      361 aaggaaatgg actccttaag attcttgtac gacggtatta gaattcaagc tgatcagacc
      421 cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga acagattggt
      481 ggcgaagtgc gtaccattaa agtttttacc accgtggata acatcaacct gcatacccag
      541 gttgttgata tgagcatgac ctatggtcag cagtttggtc cgacctatct ggatggtgca
      601 gatgttacca aaatcaaacc gcataatagc catgaaggca aaacctttta tgtgctgccg
      661 aatgatgata ccctgcgtgt tgaagcattt gagtattatc ataccaccga tccgagcttt
      721 ctgggtcgtt atatgagcgc actgaatcat accaaaaaat ggaaatatcc gcaggttaat
      781 ggtctgacca gcattaaatg ggcagataat aactgttatc tggcaaccgc actgctgaca
      841 ctgcagcaga ttgaactgaa atttaaccct ccggcactgc aggatgccta ttatcgtgca
      901 cgtgccggtg aagcagcaaa tttttgtgca ctgattctgg cctattgcaa caaaaccgtt
      961 ggtgaactgg gtgatgttcg tgaaaccatg agctacctgt ttcagcatgc aaatctggat
     1021 agctgtaaac gtgttctgaa tgttgtttgt aaaacctgcg gtcagcagca gaccacactg
     1081 aaaggtgtgg aagcagttat gtatatgggc accctgagct atgaacagtt caaaaaaggt
     1141 gttcagattc cgtgtacctg tggtaaacag gcaaccaaat atctggttca gcaagaaagc
     1201 ccgtttgtga tgatgagtgc accgcctgca cagtatgagc tgaaacatgg cacctttacc
     1261 tgtgcaagcg aatacaccgg taattatcag tgtggccatt ataaacacat caccagcaaa
     1321 gaaaccctgt attgtattga tggtgccctg ctgacaaaaa gcagcgaata taaaggtccg
     1381 attaccgacg tgttctataa agaaaatagc tacaccacca ccattaaata ataataagcg
     1441 gccgcacatc accatcacta agcttaatta gctgagcttg gactcctgtt gatagatcca
     1501 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     1561 tttattggtg agaatccaag ctagccatga acaataaaac tgtctgctta cataaacagt
     1621 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta
     1681 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1741 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1801 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1861 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcttgg
     1921 ttactcacga ctgcgatccc cggcaaaaca gcattccagg tattagaaga atatcctgat
     1981 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2041 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2101 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2161 gaacaagtct ggaaagaaat gcacaaactt ttgccattct caccggattc agtcgtcact
     2221 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2281 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2341 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2401 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt
     2461 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     2521 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     2581 cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc
     2641 tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact
     2701 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     2761 cgctctagat tacgtgcagt cgatgataag ctgtcaaaca tgagaattgt gcctaatgag
     2821 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2881 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggg
     2941 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     3001 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     3061 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     3121 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     3181 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     3241 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     3301 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     3361 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     3421 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     3481 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     3541 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     3601 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     3661 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     3721 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     3781 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     3841 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     3901 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     3961 gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc ggaatttcgg
     4021 gcagcgttgg gtcctggcca cgggtgcgca tgatctagag ctgcctcgcg cgtttcggtg
     4081 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4141 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4201 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     4261 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4321 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5161 cgaaaactca cgttaaggga ttttggtcat gacattaacc tataaaaata ggcgtatcac
     5221 gaggcccttt cgtcttcac
//
