LOCUS       4a_pET-29b(+)_NS        6117 bp DNA     circular SYN 14-APR-2023
DEFINITION  UniProtKB - P13717
            22-266.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  AUTHORS   kelvinlau
  TITLE     Direct Submission
  JOURNAL   Exported Mar 13, 2024 from SnapGene 7.1.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1144..1600
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     source          3403..3545
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    22..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)."
                     /note="color: #31849b"
     RBS             61..83
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     misc_feature    join(88..885,886..975)
                     /label=to be synth
                     /note="This feature has 2 segments:
                        1:  88 .. 885 / #a6acb3
                        2: 886 .. 975 / #a6acb3"
     CDS             91..150
                     /codon_start=1
                     /product="https://amb-express.springeropen.com/articles/10.
                     1186/s13568-017-0394-1/tables/2"
                     /label=NSP4 ss peptide
                     /note="color: #ff9900
                     Cleavage site after base 150"
                     /translation="MKKITAAAGLLLLAAQPAMA"
     CDS             151..885
                     /codon_start=1
                     /product="UniProtKB - P13717"
                     /label=Serratia marcescens nucA nuclease 22-266
                     /note="color: #ffcc00"
                     /translation="DTLESIDNCAVGCPTGGSSNVSIVRHAYTLNNNSTTKFANWVAYH
                     ITKDTPASGKTRNWKTDPALNPADTLAPADYTGANAALKVDRGHQAPLASLAGVSDWES
                     LNYLSNITPQKSDLNQGAWARLEDQERKLIDRADISSVYTVTGPLYERDMGKLPGTQKA
                     HTIPSAYWKVIFINNSPAVNHYAAFLFDQNTPKGADFCQFRVTVDEIEKRTGLIIWAGL
                     PDDVQASLKSKPGVLPELMGCKN"
     CDS             898..927
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             934..963
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             976..993
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      1060..1107
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      1144..1599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     misc_feature    1590
                     /label=differt in other pet29b
                     /note="color: #a6acb3"
     misc_feature    1600
                     /label=missing_in_other_pet29b
                     /note="color: #a6acb3"
     CDS             complement(1692..2507)
                     /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"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2629..3217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    3403..3545
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     misc_feature    3453..3455
                     /label=missing_in_other_pet29
                     /note="color: #339966"
     CDS             complement(3647..3838)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4647..5729)
                     /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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5730..5807)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
       61 tttgtttaac tttaagaagg agatatacat atgaaaaaaa ttacggcagc cgcagggttg
      121 cttcttctgg ctgctcaacc agcaatggca gacacgttag agtctattga caactgcgca
      181 gtgggttgtc caacgggagg aagctcgaat gtctcgattg tgcgtcatgc ctatacatta
      241 aacaacaact cgactacaaa gttcgccaac tgggttgctt accacatcac taaggatact
      301 cctgccagcg ggaaaacgcg taactggaaa actgacccag cattgaatcc cgcagatact
      361 cttgctccag cggactatac gggggcgaat gctgcactta aagtagaccg cggacatcag
      421 gctccgttgg cttcgttggc aggcgtaagc gactgggaga gtcttaatta ccttagcaat
      481 atcacaccgc agaaaagcga cttaaatcaa ggtgcctggg ctcgtcttga agaccaagaa
      541 cgtaaactga ttgaccgcgc tgatatttca agcgtttata cggtgacagg gcctttgtat
      601 gagcgcgata tgggaaaatt accagggaca cagaaggcgc acaccatccc cagcgcctat
      661 tggaaagtga ttttcattaa taactctcca gctgtaaacc actatgcggc atttttattt
      721 gaccagaaca cgcctaaagg cgccgatttt tgccaattcc gcgtgactgt tgatgaaatc
      781 gaaaaacgca ccggcttaat catctgggct ggccttcccg acgacgtaca agcatcctta
      841 aaaagtaagc cgggagtact tcccgaattg atgggttgta agaattctgg taacggagag
      901 caaaagctga tctctgaaga agacttaggg tctgagcaga agttaattag tgaagaggac
      961 ttataatagc tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
     1021 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     1081 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     1141 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1201 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1261 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1321 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1381 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1441 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1501 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1561 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact tttcggggaa
     1621 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1681 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1741 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1801 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1861 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1921 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     1981 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     2041 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     2101 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     2161 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     2221 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     2281 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     2341 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     2401 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2461 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta
     2521 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt
     2581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2761 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3421 ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc
     3481 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3541 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3601 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3661 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3721 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3781 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3841 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3901 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     3961 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4021 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     4081 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     4141 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     4201 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4261 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     4321 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     4381 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     4441 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     4501 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     4561 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     4621 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4681 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     4741 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     4801 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     4861 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     4921 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     4981 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     5041 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     5101 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     5161 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     5221 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     5281 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     5341 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     5401 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     5461 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     5521 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     5581 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     5641 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     5701 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     5761 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     5821 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     5881 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     5941 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     6001 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     6061 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaa
//
