LOCUS       sequence-221550-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty expression vector for 10His-SUMO tag protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  TITLE     the protein designer's laboratory toolkit
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7647)
  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            "Unpublished"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: pCDB24
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..69
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     153..172
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        153..171
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    172..196
                     /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             211..233
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             247..276
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(631..649)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      645..692
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(867..872,873..889,890..895))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(922..940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1008..1112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1113..1181,1182..1973)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1331..1350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2144..2732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2633..2652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2886..2903
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2918..3057
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2983..3002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3143..3165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3159..3350)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3986..4789)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4790..5004)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     misc_feature    5032..5534
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(6331..7413)
                     /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     7375..7394
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7414..7491)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
       61 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
      121 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
      181 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat
      241 atgggtcacc accaccacca ccaccaccac caccacggta gcctgcaaga ttcggaagtc
      301 aaccaagaag caaagccgga agtcaagccg gaagtgaaac cggaaaccca tattaacctg
      361 aaagttagtg acggcagctc tgaaattttc tttaagatca aaaagaccac gccgctgcgt
      421 cgcctgatgg aagcgtttgc caaacgtcag ggcaaggaaa tggatagcct gcgtttcctg
      481 tatgacggta ttcgcatcca ggcagatcaa gcgccggaag acctggacat ggaagacaac
      541 gacatcattg aagcccaccg tgaacagatt ggcggctcga ggatccggct gctaacaaag
      601 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      661 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
      721 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
      781 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
      841 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
      901 agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
      961 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1081 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1141 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1201 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1261 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1321 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1381 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1441 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1501 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1561 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1621 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     1681 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1741 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1801 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1861 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1921 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1981 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2041 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2101 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2161 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2221 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2281 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2341 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2401 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2461 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2521 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2581 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2641 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2701 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2761 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2821 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2881 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     2941 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     3001 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3061 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3121 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3181 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     3241 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     3301 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     3361 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     3421 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     3481 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     3541 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     3601 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     3661 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     3721 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     3781 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     3841 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     3901 gtttgcgggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     3961 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     4021 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     4081 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     4141 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     4201 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     4261 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     4321 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     4381 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     4441 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     4501 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     4561 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     4621 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     4681 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     4741 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     4801 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     4861 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     4921 ccttctgttc ggagattacc gaatcaaaaa atttcaagga aaccgaaatc aaaaaaaaga
     4981 ataaaaaaaa aatgatgaat tgaaaagcta gcttatcgat gggtcctttt catcacgtgc
     5041 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     5101 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     5161 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     5221 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     5281 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     5341 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt cttcattccg
     5401 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     5461 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     5521 acaggcaagc gatctctaag aaaccattat tatcatgaca ttaacctata aaaacgcatt
     5581 cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag
     5641 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
     5701 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
     5761 gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta
     5821 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
     5881 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
     5941 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
     6001 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     6061 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     6121 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     6181 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     6241 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     6301 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6361 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6421 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     6481 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     6541 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     6601 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     6661 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     6721 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     6781 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     6841 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     6901 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     6961 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     7021 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     7081 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     7141 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     7201 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     7261 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     7321 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     7381 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     7441 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     7501 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     7561 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     7621 ccacggggcc tgccaccata cccacgc
//
