LOCUS       sequence-217243-        4751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as TEV cleavable 
            N-terminual 8His and Sumo fusions with optional C-terminal 
            StrepII-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4751)
  AUTHORS   Lulla A, Hyvonen M
  TITLE     Protein expressionists toolkit
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: pExp-Sumo
FEATURES             Location/Qualifiers
     source          1..4751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    19..43
                     /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             56..78
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..115
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             468..491
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(505..523)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(506..523)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(581..599)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      595..642
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(878..899)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1049..2131)
                     /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     2093..2112
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2132..2209)
                     /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."
     primer_bind     2348..2367
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2521..2540)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2640..2662
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2700..2718)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2786..2890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2891..2959,2960..3751)
                     /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     complement(3109..3128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3922..4510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4411..4430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4664..4681
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa ataattttgt
       61 ttaactttaa gaaggagata tatccatgaa tcaccatcac catcaccatc accattctgg
      121 cactagtggc agcgatagcg aagtgaacca ggaagcgaaa ccggaagtta aacctgaagt
      181 gaagccggaa acccatatta acctgaaagt gtctgatggc agcagcgaaa tcttcttcaa
      241 aatcaaaaaa accactccgc tgcgtcgtct gatggaagcg tttgcgaaac gtcagggcaa
      301 agaaatggat agcctgcgtt ttctgtatga tggcattcgt attcaggcgg atcagactcc
      361 ggaagatctg gatatggaag ataacgatat tatcgaagcg catcgtgagc agattggtgg
      421 taccgaaaac ctgtacttcc agtgagacct taattaactc gagcgcatgg agccacccgc
      481 agttcgaaaa ataagcttga gtattctata gtgtcaccta aatcccagct tgatccggct
      541 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      601 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      661 tccggataac ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      721 gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
      781 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
      841 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
      901 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
      961 acagcttgca tgcctgcagg tcggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1021 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1081 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1141 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     1201 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     1261 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     1321 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     1381 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     1441 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     1501 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     1561 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     1621 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     1681 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     1741 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     1801 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     1861 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     1921 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     1981 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     2041 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     2101 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     2161 ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg gcgagctcga
     2221 attgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2281 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2341 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2401 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcctgat
     2461 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     2521 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     2581 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2641 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2701 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2761 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2821 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2881 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2941 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3001 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3061 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3121 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3181 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3241 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3301 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3361 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3421 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3481 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3541 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3601 gcgtggttct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3661 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3721 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3781 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3841 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3901 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3961 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4021 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4081 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4141 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4201 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4261 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4321 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4381 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4441 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4501 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4561 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4621 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4681 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4741 atgcagctga t
//
